Nikon ECLIPSE Ni-E Instruction Manual Download Page 62 | Manualshive

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Chapter 2

Microscopy Operations

48

Ep
i-fluo

resce

n

ce

Micro

sco

p

y

Preparation Focusing

Microscopy

Epi-fluorescence

Microscopy Procedure

1

2

3

4

5

6

7

8

9 10 11 12 13 14

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■

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Concurrent Use of Epi-Fluorescence/Phase Contrast Microscopy

By bringing the following optical elements into the optical
path, epi-fluorescence and phase contrast can be
simultaneously observed by microscope:
Filter cube for the epi-fluorescence cube turret
Ph objective
PH module for the condenser

•

If the diascopic image is not shown under

epi-fluorescence microscopy, press the
dia-illumination ON/OFF switch to turn on the lamp.

•

Remove the GIF filter if it is installed on the field

lens.

IN

OUT

1

2

A

λ

F. STO

P

A

EX.

2

3

4

5

6

Concurrent use of epi-fluorescence/phase contrast

microscopy

•

Adjust the fluorescent image brightness with ND

filters in the epi-fluorescence attachment, and
phase contrast image brightness using ND filters in
the main body. Dia-illumination should be
sufficiently dimmed using the ND filter.

ND4

ND8

ND16

Brightness adjustment of fluorescent image

ND

8

ND

32

OUT

IN

NCB

11

Adjusting brightness of phase contrast image

Ph objective

Filter cube

PH module

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Summary of Contents for ECLIPSE Ni-E

Page 1: ...Components Safety Precautions Notes on Handling the Product Individual Operations Phase Contrast Microscopy Differential Interference Contrast Microscopy Epi fluorescence Microscopy Upright Motorized Microscope Instruction Manual Operation M571 E 11 8 NF 1 1 2 M571EN01 ...

Page 2: ......

Page 3: ...nsure the accuracy of this manual errors or inconsistencies may remain If you note any points that are unclear or incorrect please contact your nearest Nikon representative Some of the equipment described in this manual may not be included in the set you have purchased If you intend to use any other equipment with this product read the manual for that equipment too If this equipment is used in a m...

Page 4: ...ssembly Maintenance Assembly Troubleshooting Maintenance and Storage Specifications Before reading the Assembly Maintenance manual read the Safety Precautions in the Operation manual Symbols Used in This Manual The following symbols are used in this manual Symbols for Safety WARNING CAUTION Highlights important information that should be noted for safety Read Safety Precautions for details Other S...

Page 5: ...he Manual Symbols Used in This Manual Safety Precautions Notes on Handling the Product Components System Configuration and Controls for Each Microscopy Before Microscopy Information Setting for Motorized Operation Operation Flowchart Bright Dark field Microscopy Epi fluorescence Microscopy Epi fluorescence Epi fluorescence Differential Interference Contrast Epi fluorescence Phase Contrast Differen...

Page 6: ...rations 11 1 Before Microscopy 12 2 Operation Flowchart 14 3 Bright Dark field Microscopy Procedure 18 4 Epi fluorescence Microscopy Procedure 33 5 Differential Interference Contrast Microscopy Procedure 49 6 Phase Contrast Microscopy Procedure 67 Individual Operations 83 1 Display Panel Details 84 2 Using Operation Buttons on Ni E 87 2 1 Front Operation Button 87 2 2 Right Operation Button 88 2 3...

Page 7: ...e Turret Operation Switching Excitation Methods 118 16 2 Selecting Filters 120 16 3 Protecting the Sample and Preventing It from Decoloration Using the Shutter 122 16 4 Adjusting the Brightness of the Fluorescent Image Using ND Filters and the Aperture Diaphragm 124 16 5 Restricting the Illumination to the Area of the Specimen being Viewed Centering and Adjusting the Field Diaphragm 126 16 6 Chang...

Page 8: ... caution at all times during use Read the relevant instructions in this manual before attempting to use or adjust any part to which the symbol has been affixed Biohazard This symbol is affixed to the front of the stand of this product to call your attention to the following WARNING Using this product may constitute a biohazard risk if a sample comes into contact with this product To avoid biohazar...

Page 9: ...ng because of its characteristics Be sure to refer to the manual for the light source being used 3 Notes on the power cord Be sure to use the specified power cord Use of other power cords may result in malfunction or fire This product is classified as having Class I protection against electric shock Make sure this product is connected to an appropriate protective earth terminal See Chapter 4 2 Per...

Page 10: ...path Removing the D UV slider from the optical path will cause your eyes to become exposed to ultraviolet light High pressure gas The lamps contain sealed gas under very high pressure And the pressure increases when the lamp is on If the lamp is scratched fouled subjected to high external pressure or physical impact or used beyond its service life the sealed gas may leak or the lamp may burst resu...

Page 11: ... injuries during operation 5 Avoid contact with water or chemical solutions Never expose this product to water or chemical solutions and avoid using this product in circumstances where there is risk of exposure to water or chemical solutions Exposure of electric parts such as the HG Precentered Fiber Illuminator to liquids may cause a short circuit resulting in malfunction or abnormal heating If w...

Page 12: ...ons Installing this product in a hot humid location may result in the formation of mold or condensation on lenses impairing performance or causing malfunctions Operating conditions temperature 0 to 40 C humidity 60 RH max no condensation Transporting storage conditions temperature 20 to 60 C humidity 90 RH max no condensation See Chapter 3 4 Transportation Using the fastening position mode switch ...

Page 13: ...er the objective as extraneous light If possible switch off the room lights directly above this product when making observations Select a location with minimal dust To avoid splashes do not use this product near water Make sure the ambient temperature is 0 to 40 C and humidity is 60 or less When transporting or storing this product the ambient temperature must be 20 to 60 C with the humidity at 90...

Page 14: ...npacking and unlocking 1 Check the package contents ECLIPSE Ni E Main Body X1 Hex driver X3 Hex wrench X2 Toolbox X1 Accessory sticker X2 types A blinder for the mounting hole for accessory not in use on the top of the arm and nameplate for an additional ND filter to the ND filter cassette 2 Before turning on the power remove the tightening screw from the front of the elevating section See Chapter...

Page 15: ...croscopy with name of components and controls The figures are examples of basic motorized systems Manually operated devices are also available For the list of components used for the ECLIPSE Ni E microscope see Chapter 1 2 Components List in the separately provided Assembly Maintenance instruction manual ...

Page 16: ...e Model NI ND6 E or NI N7 E Substage Model NI SSR NI E right handle rotatable ceramic coated stage Model NIE CSRR2 Motorized universal condenser Model NI CUD E Ni E main body Precentered lamphouse 12V 100W Model NI LH Halogen lamp 100W Model PHILIPS 7724 or OSRAM HLX64623 Motorized DSC zooming port for quadrocular tube Model NI RPZ E Camera head Contact arm Model NI CAM Connector box Model NIE CB ...

Page 17: ...arse focus knob Condenser centering screw Right control buttons See 1 3 Buttons on the next page Front control buttons See 1 3 Buttons on the next page Left control buttons See 1 3 Buttons on the next page Stage X knob Coarse focus knob Moving claw of the specimen holder Stage Y knob Fine focus knob Condenser focus knob Condenser centering screw Stage rotation clamp screw Fine focus knob ...

Page 18: ...l brightness adjustment button DISPLAY Previous button BINO optical path button Power LED FRONT optical path button Switch DISPLAY button ND filter IN OUT switch FL CUBE CCW rotation button Factory setting First epi fluorescence cube turret DIA field diaphragm button FL SHUTTER button Factory setting Shutter built into first epi fluorescence cube turret CAPTURE button Factory setting Camera direct...

Page 19: ...CC Rules Operation is subject to the following two conditions 1 this device may not cause harmful interference and 2 this device must accept any interference received including interference that may cause undesired operation This Class A digital apparatus complies with Canadian ICES 003 Cet appareil numérique de la classe A est confirme à la norme NMB 003 du Canada 5 4 0 0 0 1 Control Box A Front ...

Page 20: ...k field Microscopy Components IN ND 8 ND 32 NCB 11 OUT 1 2 A λ F STOP A STOP EX ADJ 2 3 4 5 6 INTENSILIGHT C HGFIE LAMP POWER RUN TIME hrs 50 60 70 See the figure for bright field system for control box A and DS L3 DS Camera Control Unit Motorized epi fluorescence cube turret Model NI FLT6 E Epi fluorescence attachment Model NI FLEI HG adapter Model C HGFIB Light shielding plate Motorized HG prece...

Page 21: ...ND8 ND16 50 60 70 ND filter slider FL DIC analyzer slider NI FA used for epi fluorescence differential interference contrast microscopy Filter cube replacement slot cover Field diaphragm lever Aperture diaphragm lever Field diaphragm centering screws Aperture diaphragm centering screws Filter cube name plate window ...

Page 22: ...py IN ND 8 ND 32 NCB 11 OUT 1 2 A λ 50 60 70 A DIC module must be installed on the motorized universal condenser Epi fluorescence components are also required when performing differential interference contrast epi fluorescence microscopy See the figure for bright field system for control box A and DS L3 DS Camera Control Unit Motorized nosepiece Model NI ND6 E DIC rotating polarizer D DP DIC objec...

Page 23: ...he left rear side A PH module must be installed on the motorized universal condenser Keep the PH module centering knob of the motorized universal condenser facing down during microscopy Lift the knob only when centering and push it into the centering screw to use Epi fluorescence components are also required for concurrent use of phase contrast epi fluorescence microscopy See the figure for bright...

Page 24: ...Chapter 1 Components 10 Components ...

Page 25: ...on 4 Phase contrast microscopy The procedures described herein assume that all required components are attached to the microscope with all necessary cables properly connected and that information registration for motorized operation has been completed The function of operation buttons of the main body has been set to the default When using the microscope for the first time or changing the motorize...

Page 26: ...ation Filter Barrier Filter Information CONNECTION Chapter 3 23 1 2 Destination camera to output signal from the DSC connector Connector DSC1 Rear of the microscope Connector DSC2 Connector box Not connected DS L3 Chapter 6 3 1 Configuring the Connection of Digital Camera Motorized shutter connected to a SHUTTER connector Connector EPI SHUTTER Rear of the microscope Connector DIA SHUTTER Connector...

Page 27: ...e Elevating Section Rotation stop of the nosepiece depending on the position of the elevating section No rotation stop DS L3 Chapter 6 5 3 Disabling the Rotation of the Motorized Nosepiece Depending on the Position of the Elevating Section Reverse rotation stop of the nosepiece No rotation stop DS L3 Chapter 6 5 4 Disabling the Reverse Rotation of the Nosepiece Toggle rotation of the nosepiece Tog...

Page 28: ...opy Operations 14 Before Microscopy Operation Flowchart 2 Operation Flowchart The operation flowchart for a microscopy is shown below See the procedure for each microscopy in the next and subsequent sections for details ...

Page 29: ...Chapter 2 Microscopy Operations 15 Before Microscopy Operation Flowchart ...

Page 30: ...Chapter 2 Microscopy Operations 16 Before Microscopy Operation Flowchart ...

Page 31: ...Chapter 2 Microscopy Operations 17 Before Microscopy Operation Flowchart ...

Page 32: ...n the MICROSCOPE CONTROL screen can be changed as desired For details on the procedure for controlling the microscope from DS L3 see Chapter 4 Microscopy with DS L3 in the DS L3 instruction manual Microscope Operation Also see Chapter 3 23 Operation on DS L3 23 2 Controlling the Microscope in this manual Preparation 1 Turn on the power Press the power switch to to turn on the power 1 Turn on the p...

Page 33: ...play pattern 1 Factory setting example 2 Turn on the dia illumination lamp Press the dia illumination lamp ON OFF switch Turning on dia illumination lamp PA__2x____Z _____0 000um DIA ON__________FS30 0mm Voltage is not displayed Turning lamp ON brightness adjustment control The lamp can also be turned on and adjusted from DS L3 by configuring the lamp control button on the DS L3 s MICROSCOPE CONTR...

Page 34: ...r FRONT optical path button 100 to tube adapter REAR optical path button 100 to rear port Binocular 100 with BINO button PA__2x____Z _____0 000um BINO____ZM 1 0x_FS30 0mm BINO button Binocular optical path 100 4 Lower the condenser slightly from the uppermost position Turn the condenser focus knob until the condenser is positioned at the upper limit where it clicks to a stop and then lower it a li...

Page 35: ...ragm Press the left side mark side of the DIA field diaphragm and DIA aperture diaphragm button Pressing the right side mark side of both buttons will close the diaphragm ND 8 ND 32 OUT IN NCB 11 Fully opening the field and aperture diaphragms PA__2x____Z _____0 000um BF____AS30 6mm__FS30 6mm F STOP button sub screen DIA field diaphragm open A STOP button sub screen DIA aperture diaphragm open DIA...

Page 36: ...ht field position Press the FUNCTION button 1 2 to turn the condenser turret and display OPEN on the front panel of Ni E The turret s empty position enters the optical path in this state Button 1 is for counterclockwise rotation and button 2 is for clockwise rotation IN ND 8 ND 32 NCB 11 OUT Moving the condenser to the empty position PA__2x____Z _____0 000um OPEN__AS30 0mm__FS30 0mm CONDEN button ...

Page 37: ...adjacent address 6 7 Intelligent nosepiece Manually switch the objective since motorized operation is not available for the intelligent nosepiece On DS L3 only information display is provided Bringing 10x in optical path using the OBJ CW CCW button PA_10x____Z _____0 000um OPEN__AS30 0mm__FS30 0mm NOSEPIECE button sub screen 10x 8 Place a specimen on the stage and move the stage to bring the targe...

Page 38: ...ing Procedure Z RESET The Z axis coordinate value can be reset to zero This is useful when you wish to use the current position as a reference in adjusting the focus See Chapter 3 4 3 Resetting the Z axis Coordinate Refocusing This operation moves the Z axis position to the retracting position or returns it to the original position See Chapter 3 4 4 Refocusing Brightness adjustment Brightness adju...

Page 39: ...asy adjustment look into the eyepiece as if you were looking at a distant object 50 60 7 0 Adjusting the interpupillary distance 12 Focus and center the condenser 1 Press the DIA field diaphragm button to close the field diaphragm to a minimum and look into the eyepiece Focus on the field diaphragm image using the condenser focus knob and then adjust the condenser centering screws with a tool to c...

Page 40: ...isplay DF on the front panel of Ni E The turret s dark field module enters the optical path in this state Button 1 is for counterclockwise rotation and button 2 is for clockwise rotation IN ND 8 ND 32 NCB 11 OUT Bring the dark field module into the optical path PA__2x____Z _____0 000um DF____AS30 0mm__FS30 0mm Dark field module The dark field module can be used with any objective with a numerical ...

Page 41: ...een the specimen and the objective See Chapter 3 14 Oil Water Immersion Never apply oil between the universal condenser top lens and the specimen The universal condenser is designed for dry use Note that an oil water immersion type objective cannot be used for the motorized universal condenser Motorized nosepiece toggling Two objectives can be alternated by specifying an objective pair Objectives ...

Page 42: ...gm opening will degrade the image resolution Nikon does not recommend setting the aperture diaphragm to less than 60 of the numerical aperture of the objective Adjustment timing for the aperture diaphragm Be sure to adjust the aperture diaphragm each time you change the objective Î For dark field microscopy the aperture diaphragm is not adjusted here Keep it open ND 8 ND 32 OUT IN NCB 11 Adjusting...

Page 43: ...t circumscribes the field of view Excessively opening the field diaphragm will result in stray light entering the field of view generating flare and reducing the image contrast It will also cause the specimen to become decolorized over a wider area Adjustment timing for the field diaphragm Be sure to adjust the field diaphragm each time you change the objective ND 8 ND 32 OUT IN NCB 11 Adjusting f...

Page 44: ...etting for improved color reproduction Changing the lamp voltage for brightness adjustment will affect the lamp color temperature and alter the color balance of the image Increasing the voltage will result in a bluish tint while reducing the voltage results in a reddish tint If accurate color reproduction is critical set the dia illumination lamp brightness control knob at the mark to adjust the l...

Page 45: ...r look away from the eyepiece during the switchover Low magnification microscopy If you are using a universal condenser during bright field microscopy with a 2x or 4x objective attach the 2 4x auxiliary lens to the turret and move it into the optical path When doing so fully open the aperture and field diaphragms for the maximum range of illumination NI FA FL DIC Analyzer Slider Dummy Slider When ...

Page 46: ...field Microscopy 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 19 Turn off the power Turn off the power switches press to the O position on the control box A and connected motorized devices Each power LED turns off 50 60 70 Turning off the device Power switch OFF Power LED OFF Power LED OFF Power LED OFF Power switch OFF Power LED OFF ...

Page 47: ... are shown for procedures that can be performed from DS L3 The content and layout of buttons on the screens in the figure and the actual MICROSCOPE CONTROL screen may differ because the setting of buttons on the MICROSCOPE CONTROL screen can be changed as desired For details on the control from DS L3 see Chapter 3 23 Operation on DS L3 23 2 Controlling the Microscope in this manual WARNING The lig...

Page 48: ... habit of performing this operation ND 8 ND 32 OUT IN NCB 11 Closing shutter PF__2x____Z _____0 000um UV 2B C____ND_1 C__EPI C C Shutter closed Shutter blocking epi fluorescence light The shutter is built into the epi fluorescence cube turret and is also equipped on the HG precentered fiber illuminator Also an EPI motorized shutter can be attached to the microscope Epi fluorescence illumination ca...

Page 49: ...onents for the characteristics of the specimen and the fluorescence dye See Chapter 3 16 2 Selecting Filters ND 8 ND 32 OUT IN NCB 11 Bringing the filter cube into the optical path PF_10x____Z _____0 000um UV 2B C____ND_1 C__EPI C Intelligent epi fluorescence cube turret Switch the intelligent epi fluorescence cube turret manually because it is not motorized On DS L3 only information display is pr...

Page 50: ...n the clockwise direction as viewed from above while the OBJ CCW button turns the nosepiece in the counterclockwise direction Oil immersion When using an oil immersion type objective apply Nikon specified non fluorescence immersion oil between the specimen and the objective See Chapter 3 14 Oil Water Immersion Motorized nosepiece toggling Two objectives can be alternated by specifying an objective...

Page 51: ...in the motorized epi fluorescence cube turret Other shutters If the shutter of the HG precentered fiber illuminator or EPI motorized shutter is closed be sure to open these shutters as well Label of the button changes indicating the current open close status OPEN open CLOSED closed When you are using intelligent epi fluorescence cube turret the SHUTTER FL button does not appear Therefore it is not...

Page 52: ...void the following action which can cause equipment malfunction Rotating the right and left focus knobs in opposite directions Do not turn the focus knobs in opposite direction 9 Center the field diaphragm of the epi fluorescence attachment 1 Use the field diaphragm lever on the epi fluorescence attachment to close the field diaphragm and then move the field diaphragm image to the center of the fi...

Page 53: ...lare and reducing the image contrast In addition the sample will become decolorized over a wider area Field diaphragm s adjustment timing Be sure to adjust the field diaphragm each time you change the objective Appropriate size of the field diaphragm Adjusting the field diaphragm 11 Adjust the brightness Use ND filters on the epi fluorescence attachment to adjust image brightness The aperture diap...

Page 54: ...e disappear Bright ambient lights will make it difficult to view the image Nikon recommends keeping the room dark during fluorescence observations NI FA FL DIC Analyzer Slider Dummy Slider In the epi fluorescence microscopy the FL DIC analyzer slider inserted in the epi fluorescence cube turret is not used When the analyzer slider is inserted in the epi fluorescence cube turret pull it out to the ...

Page 55: ...ure 1 2 3 4 5 6 7 8 9 10 11 12 13 14 14 Turn off the power Turn off the power switches press to the O position on the control box A and connected motorized devices Each power LED turns off 5 6 1 A λ 5 4 3 2 1 50 60 70 Turning off the device Power switch OFF Power LED OFF Power LED OFF Power LED OFF Power switch OFF Power LED OFF ...

Page 56: ...d when the FL SHUTTER button is lit ND 8 ND 32 OUT IN NCB 11 Closing shutter 2 Rotate the epi fluorescence cube turret to bring turret address 1 into the optical path If a filter cube is in address 1 remove it in advance ND 8 ND 32 OUT IN NCB 11 Removing the filter cube PF__2x____Z _____0 000um OPEN C_____ND_1 C__EPI C Empty position Shutter closed 3 Press the dia illumination ON OFF switch to tur...

Page 57: ...ination off Check that the epi fluorescence illumination path shutter is closed The shutter is closed when the FL SHUTTER button is lit ND 8 ND 32 OUT IN NCB 11 Closing shutter 2 All optical elements required for differential interference contrast microscopy are removed from the optical path in procedures 6 and 7 in 5 Differential Interference Contrast Microscopy Procedure At the same time Remove ...

Page 58: ...ret to bring the desired filter cube into the optical path ND 8 ND 32 OUT IN NCB 11 Bringing the filter cube into the optical path Remove the FL DIC analyzer slider from optical path A F STOP 2 3 4 5 6 50 60 70 Removing the FL DIC analyzer slider from optical path Fully open the field diaphragm and aperture diaphragm of the epi fluorescence attachment F STOP A STOP EX ADJ Fully opening field and a...

Page 59: ...sition in the DIC analyzer slider the analyzer is brought into the optical path It is removed from the optical path when the slider is pulled back to the first click stop position IN OUT 1 2 A λ F STOP A EX 2 3 4 5 6 Concurrent use of epi fluorescence differential interference contrast microscopy ND4 ND8 ND16 Brightness adjustment of fluorescent image If the diascopic image is not shown under epi ...

Page 60: ...red fiber illuminator for epi fluorescence illumination off Check that the epi fluorescence illumination path shutter is closed The shutter is closed when the FL SHUTTER button is lit ND 8 ND 32 OUT IN NCB 11 Closing shutter 2 The position of the condenser turret is set to OPEN bright field position in procedure 6 in 6 Phase Contrast Microscopy Procedure but Remove the filter cube from the optical...

Page 61: ... cube into the optical path ND 8 ND 32 OUT IN NCB 11 Bringing the filter cube into the optical path Fully open the field diaphragm and aperture diaphragm of the epi fluorescence attachment F STOP A STOP EX ADJ Fully opening field and aperture diaphragms 4 Turn on the power of the HG precentered fiber illuminator and continue from procedure 6 in the epi fluorescence microscopy procedure Aperture di...

Page 62: ... diascopic image is not shown under epi fluorescence microscopy press the dia illumination ON OFF switch to turn on the lamp Remove the GIF filter if it is installed on the field lens IN OUT 1 2 A λ F STOP A EX 2 3 4 5 6 Concurrent use of epi fluorescence phase contrast microscopy Adjust the fluorescent image brightness with ND filters in the epi fluorescence attachment and phase contrast image br...

Page 63: ...ay differ because the setting of buttons on the MICROSCOPE CONTROL screen can be changed as desired For details on the control from DS L3 see Chapter 3 23 Operation on DS L3 23 2 Controlling the Microscope in this manual Preparation 1 Turn on the power Press the power switch to to turn on the power 1 Turn on the power switches for all accessory devices connected Excluding DS L3 The power LED on ea...

Page 64: ...lay Display pattern 1 Factory setting example 2 Turn on the dia illumination lamp Press the dia illumination lamp ON OFF switch Turning on the dia illumination lamp and adjust brightness PA__2x____Z _____0 000um DIA ON__________FS30 0mm Voltage is not displayed Turning lamp ON brightness adjustment control The lamp can also be turned on and adjusted from DS L3 by configuring the lamp control butto...

Page 65: ... button 100 to binocular FRONT optical path button 100 to tube adapter REAR optical path button 100 to rear port Binocular 100 with BINO button PA__2x____Z _____0 000um BINO____ZM 1 0x_FS30 0mm BINO button Binocular optical path 100 4 Lower the condenser slightly from the uppermost position Turn the condenser focus knob until the condenser is positioned at the upper limit where it clicks to a stop...

Page 66: ...ragm and aperture diaphragm Press the left side mark side of the DIA field diaphragm and DIA aperture diaphragm button Pressing the right side mark side of both buttons will close the diaphragm ND 8 ND 32 OUT IN NCB 11 Fully opening the field and aperture diaphragms PA__2x____Z _____0 000um OPEN__AS30 6mm__FS30 6mm F STOP button sub screen DIA field diaphragm open A STOP button sub screen DIA aper...

Page 67: ... to the OPEN empty bright field position Press the FUNCTION button 1 2 to turn the condenser turret and display OPEN on the front panel of Ni E The turret s empty position enters the optical path in this state Button 1 is for counterclockwise rotation and button 2 is for clockwise rotation IN ND 8 ND 32 NCB 11 OUT Moving the condenser to the empty position PA__2x____Z _____0 000um OPEN__AS30 0mm__...

Page 68: ... OBJ CW or CCW button to bring the 10x objective into the optical path Pressing the OBJ CW button turns the nosepiece by one address in the clockwise direction as viewed from above while the OBJ CCW button turns the nosepiece in the counterclockwise direction Restrict movement of motorized nosepiece A function to prevent the objective from touching the specimen when the motorized nosepiece is rota...

Page 69: ...X Y knobs to move the stage and bring the target into the optical path See Chapter 3 5 Bringing the Target into the Optical Path Horizontal Stage Movement Rotation So that the sample sealed under the cover glass will be lighted Non fluorescent distortion free slide glass Use clean non fluorescent distortion free slides and cover glasses Setting specimen Motorized horizontal stage movement When usi...

Page 70: ... 4 1 Proper Focusing Procedure Z RESET The Z axis coordinate value can be reset to zero This is useful when you wish to use the current position as the reference position in adjusting the focus See Chapter 3 4 3 Resetting the Z axis Coordinate Refocusing This operation moves the Z axis position to the retracting position or returns it to the original position See Chapter 3 4 4 Refocusing Brightnes...

Page 71: ...a distant object 50 60 7 0 Adjusting the interpupillary distance 13 Focus and center the condenser 1 Press the DIA field diaphragm button to close the field diaphragm to a minimum and look into the eyepiece Focus on the field diaphragm image using the condenser focus knob and then adjust the condenser centering screws with a tool to center the diaphragm image within the field of view 2 Operate the...

Page 72: ...the optical path 5 Loosen the polarizer rotation screw to rotate the polarizer align the line with to the symbol and then tighten the rotation clamp screw 6 Rotate the stage knob to move the specimen and bring a portion where there is no sample under the cover glass into the optical path 7 Adjust the attaching angle of the polarizer to the microscope Perform by one of the following methods Method ...

Page 73: ... condenser is designed for dry use Note that an oil water immersion type objective cannot be used for the motorized universal condenser Motorized nosepiece toggling Two objectives can be alternated by specifying an objective pair Objectives can be switched using the OBJ CW CCW button on the microscope See Chapter 3 11 1 Motorized Nosepiece Operation Bringing desired DIC objective into optical path...

Page 74: ...jective into the optical path Button 1 is for counterclockwise rotation and button 2 is for clockwise rotation Display DIC module N1 etc on the NI E s display panel The turret s DIC module enters the optical path in this state Choose the DIC module appropriate for the intended microscopy purpose and the objective used See Chapter 3 17 Differential Interference Contrast Microscopy 17 2 Using Optica...

Page 75: ...ture of the objective Since an excessively small aperture diaphragm opening will degrade the image resolution Nikon does not recommend setting the aperture diaphragm to less than 60 of the numerical aperture of the objective Adjustment timing for the aperture diaphragm Be sure to adjust the aperture diaphragm each time you change the objective ND 8 ND 32 OUT IN NCB 11 Adjusting aperture diaphragm ...

Page 76: ... be adjusted so that it circumscribes the field of view Excessively opening the field diaphragm will result in stray light entering the field of view generating flare and reducing the image contrast It will also cause the specimen to become decolorized over a wider area Adjustment timing for the field diaphragm Be sure to adjust the field diaphragm each time you change the objective ND 8 ND 32 OUT...

Page 77: ...tomicrography voltage setting for improved color reproduction Changing the lamp voltage for brightness adjustment will affect the lamp color temperature and alter the color balance of the image Increasing the voltage will result in a bluish tint while reducing the voltage results in a reddish tint If accurate color reproduction is critical set the dia illumination lamp brightness control knob at t...

Page 78: ...ng direction Microscopy with color contrast Placing the NCB filter into the optical path and inserting the D LP lambda plate into the lambda plate slot on the DIC nosepiece will turn the background of the field of view to a sensitive color enabling observations with high color contrast An interference color is displayed based on changes in the refraction index and or thickness of the specimen NI F...

Page 79: ...trast Microscopy 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 23 Turn off the power Turn off the power switches press to the O position on the control box A and connected motorized devices Each power LED turns off 5 6 1 A λ 5 4 3 2 1 50 60 70 Turning off the device Power LED OFF Power LED OFF Power switch OFF Power LED OFF Power switch OFF Power LED OFF ...

Page 80: ... JAPAN D D P Polarizer removed from optical path 1 2 A λ 10x Removing the optical element for differential interference contrast microscopy from the optical path 2 Operate the motorized universal condenser to bring the OPEN position into the optical path PA__2x____Z __1235 555um OPEN__AS12 3mm__FS25 5mm Low magnification microscopy To perform bright field microscopy with a 2x or 4x objective attac...

Page 81: ...ting of buttons on the MICROSCOPE CONTROL screen can be changed as desired For details on the control from DS L3 see Chapter 3 23 Operation on DS L3 23 2 Controlling the Microscope in this manual Preparation 1 Turn on the power Press the power switch to to turn on the power 1 Turn on the power switches for all accessory devices connected Excluding DS L3 The power LED on each device will light up 2...

Page 82: ...isplay pattern 1 Factory setting example 2 Turn on the dia illumination lamp Press the dia illumination lamp ON OFF switch Turning on the dia illumination lamp PA__2x____Z _____0 000um DIA ON__________FS30 0mm Voltage is not displayed Turning lamp ON brightness adjustment control The lamp can also be turned on and adjusted from DS L3 by configuring the lamp control button on the DS L3 s MICROSCOPE...

Page 83: ...nocular FRONT optical path button 100 to tube adapter REAR optical path button 100 to rear port Binocular 100 with BINO button PA__2x____Z _____0 000um BINO____ZM 1 0x_FS30 0mm BINO button Binocular optical path 100 4 Lower the condenser slightly from the uppermost position Turn the condenser focus knob until the condenser is positioned at the upper limit where it clicks to a stop and then lower i...

Page 84: ...re diaphragm Press the left side mark side of the DIA field diaphragm and DIA aperture diaphragm button Pressing the right side mark side of both buttons will close the diaphragm ND 8 ND 32 OUT IN NCB 11 Fully opening field and aperture diaphragms PA__2x____Z _____0 000um OPEN__AS30 6mm__FS30 6mm F STOP button sub screen DIA field diaphragm open A STOP button sub screen DIA aperture diaphragm open...

Page 85: ...y bright field position Press the FUNCTION button 1 2 to turn the condenser turret and display OPEN on the front panel of Ni E The turret s empty position enters the optical path in this state Button 1 is for counterclockwise rotation and button 2 is for clockwise rotation IN ND 8 ND 32 NCB 11 OUT Moving the condenser to the empty position PA__2x____Z _____0 000um OPEN__AS30 0mm__FS30 0mm CONDEN b...

Page 86: ...ss 1 to adjacent address 6 7 Intelligent nosepiece Manually switch the objective since motorized operation is not available for the intelligent nosepiece On DS L3 only information display is provided Bringing 10x in optical path using the OBJ CW CCW button PA_10x____Z _____0 000um OPEN__AS30 0mm__FS30 0mm NOSEPIECE button sub screen 10x 8 Place a specimen on the stage and move the stage to bring t...

Page 87: ... Focusing Procedure Z RESET The Z axis coordinate value can be reset to zero This is useful when you wish to use the current position as a reference in adjusting the focus See Chapter 3 4 3 Resetting the Z axis Coordinate Refocusing This operation moves the Z axis position to the retracting position or returns it to the original position See Chapter 3 4 4 Refocusing Brightness adjustment Brightnes...

Page 88: ... For easy adjustment look into the eyepiece as if you were looking at a distant object 50 60 7 0 Adjusting the interpupillary distance 12 Focus and center the condenser 1 Press the DIA field diaphragm button to close the field diaphragm to a minimum and look into the eyepiece Focus on the field diaphragm image using the condenser focus knob and then adjust the condenser centering screws with a too...

Page 89: ...front side of the Ni E main body The PH module for the turret is brought into the optical path Button 1 is for counterclockwise rotation and button 2 is for clockwise rotation Chapter 3 18 Phase Contrast Microscopy 18 2 Using Optical Elements IN ND 8 ND 32 NCB 11 OUT Bringing the PH module Ph1 into optical path PF_10x____Z __1234 567um Ph1___AS30 0mm__FS30 0mm GIF filter The GIF filter green inter...

Page 90: ...rotate the eyepiece to focus on the phase plate of the objective 5 If the phase plate of the objective is not aligned with the PH module Ph annular diaphragm of the condenser align them by raising and pushing the two PH module centering knobs on both sides of the back of the condenser and turning the centering screws When done restore the centering knobs to the original position 6 Place the eyepie...

Page 91: ... oil between the specimen and the objective See Chapter 3 14 Oil Water Immersion Never apply oil between the universal condenser top lens and the specimen The universal condenser is designed for dry use Note that an oil water immersion type objective cannot be used for the motorized universal condenser Motorized nosepiece toggling Two objectives can be alternated by specifying an objective pair Ob...

Page 92: ...ule with the same Ph code as the objective into optical path Button 1 is for counterclockwise rotation and button 2 is for clockwise rotation Ph code One of the Ph codes Ph1 Ph2 or Ph3 is indicated on the Ph objective depending on the size of the phase plate Ph codes have nothing to do with the magnification of the objective Always use a Ph objective and PH module with the same Ph code You cannot ...

Page 93: ...ctive Since an excessively small aperture diaphragm opening will degrade the image resolution Nikon does not recommend setting the aperture diaphragm to less than 60 of the numerical aperture of the objective Adjustment timing for the aperture diaphragm Be sure to adjust the aperture diaphragm each time you change the objective ND 8 ND 32 OUT IN NCB 11 Adjusting aperture diaphragm PA_40x____Z __12...

Page 94: ...that it circumscribes the field of view Excessively opening the field diaphragm will result in stray light entering the field of view generating flare and reducing the image contrast It will also cause the specimen to become decolorized over a wider area Adjustment timing for the field diaphragm Be sure to adjust the field diaphragm each time you change the objective ND 8 ND 32 OUT IN NCB 11 Adjus...

Page 95: ...aging Photomicrography voltage value is adjusted and set at shipment and users cannot modify it To adjust the brightness further without changing color bring the NCB11 filter in the filter cassette into the optical path Use the ND filters for adjustment See Chapter 3 3 Adjusting the Brightness of a Diascopic Image Photomicrography voltage setting Viewing the specimen 21 View the specimen To observ...

Page 96: ...motorized devices Each power LED turns off 50 60 70 Turning off the device Switch from Phase Contrast Microscopy to Bright field Microscopy Operate the condenser turret to bring the OPEN position into the optical path For details on other operations and precautions see 3 Bright Dark field Microscopy Procedure Switch from Phase Contrast Microscopy to Epi Fluorescence Microscopy See the end of 4 Epi...

Page 97: ...83 Individual Operations 3 Chapter 3 Individual Operations This chapter describes how to use the display panel operation buttons and each part device of the Ni E main body in detail ...

Page 98: ...rret 1 1st layer 5 Motorized universal condenser 11 Motorized epi fluorescence cube turret 2 2nd layer 6 DIA aperture diaphragm 12 Motorized excitation filter wheel 3 Initialization completion display Either Microscope state or FUNCTION button function can be displayed on the display panel When the initialization is completed pattern 1 of the microscope state is displayed first Press the DISPLAY S...

Page 99: ...e cube turret 1st layer epi fluorescence cube turret s built in shutter 1st layer X axis position unit um Zoom magnification Y axis position unit um UV 2B C__ X ___ 0 000um ZM 1 0x___Y _____0 000um Pattern 7 Top Bottom Objective X axis position unit um Zoom magnification Y axis position unit um PA__2x___ X ___ 0 000um ZM 1 0x___Y _____0 000um Pattern 8 Top Bottom BA barrier filter status status of...

Page 100: ...cube turret UV 2B Name of the filter cube in the optical path of the motorized intelligent filter cube 2nd layer Example Motorized XY stage position X _000000 000um Motorized XY stage X coordinate Y _000000 000um Motorized XY stage Y coordinate EPI motorized shutter status EPI O EPI motorized shutter open OPEN EPI C EPI motorized shutter closed CLOSE Motorized barrier filter wheel BA000 000 Barrie...

Page 101: ...filter wheel counterclockwise and clockwise Buttons 5 and 6 Rotate the barrier filter wheel counterclockwise and clockwise 12 BINO optical path button Direct 100 of the optical path to the binocular with status indicator LED 100 to the binocular when the LED is on and 100 to a tube adapter or to a rearport when the LED is off 13 FRONT optical path button Direct 100 of the optical path to the tube ...

Page 102: ...ess counterclockwise as viewed from above 20 FL SHUTTER button Open or close the 1st layer motorized epi fluorescence cube turret built in shutter The shutter is closed when the SHUTTER button is ON 21 CAPTURE button Capture an image using a digital camera connected via USB on the rear of the main body 22 DIA aperture diaphragm button Adjust the diameter of the condenser s aperture diaphragm 23 DI...

Page 103: ...ntrol knob Adjust the brightness of the dia illumination lamp by changing the supplied voltage 29 Dia illumination ON OFF switch Turn the dia illumination lamp on or off Changing the function of an operation button For buttons with appended in the table FUNCTION button FL CUBE CW CCW button OBJ CW CCW button FL SHUTTER button and CAPTURE button their functions can be changed See 23 Operation on DS...

Page 104: ...perature and alter the color balance of the image Increasing the voltage will result in a bluish tint while reducing the voltage results in a reddish tint If accurate color reproduction is critical set the brightness control knob at the mark and bring the NCB11 filter in the filter cassette into the optical path for optimal color reproduction To adjust the brightness use the ND filters Photomicrog...

Page 105: ...st accept any interfer ence receive d includi ng interfer ence that may cause undesi red operat ion This Class A digital appara tus compli es with Canad ian ICES 0 03 Cet appare il numéri que de la classe A est confirm e à la norme NMB 0 03 du Canad a 910001 ND 1 2 3 Do not touch the lamphouse while the lamp is lit The surface of the lamphouse becomes hot when the lamp is on Turn off the power and...

Page 106: ...1 Bring the 10x objective into the optical path and turn the coarse focus knob to raise the stage to the upper limit 2 Direct 100 of the light to the binocular section 3 Adjust the brightness of dia illumination 4 Look into the eyepiece and turn the coarse focus knob to lower the stage and focus on the specimen 5 Turn the fine focus knob to further adjust the focus Focusing by the focus knob PA__2...

Page 107: ...cal correction The difference between focal points of objectives is automatically corrected when switching an objective See 19 2 Interlocking Function for conditions for interlocking and interlocked operations Interlocked operation of vertical Z stage movement speed A vertical Z stage movement speed can be interlocked when switching an objective See 19 2 Interlocking Function for details on condit...

Page 108: ... removed by inserting a hex driver 2 mm across flats or the likes into the notch in the knob Position exchange of the focus knob 4 3 Resetting the Z axis Coordinate The Z axis coordinate can be reset to zero and used as the reference point for adjusting the focus To do so adjust the focus on the sample and then press the Z RESET button The Z axis coordinate value will be reset to 0 000 um Operatio...

Page 109: ...en the objective is retracted and will be turned off otherwise When the stage is retracted the elevating section cannot be moved by turning the focus knobs ESCAPE Button used to retract restore the stage Retraction Position Range 5 mm lower from the current position 10 mm lower from the current position Lower limit of the elevating section software limit See 23 Operation on DS L3 23 1 Setting Up t...

Page 110: ... Controller and 21 Using the Joystick Controller 5 1 Operating the Rotatable Ceramic Coated Stage Notes on moving the stage Avoid the following actions which can cause equipment malfunctions Moving the stage to the right and left by holding the top surface of the stage directly Forcing the stage to move or rotate past the X Y stroke or the maximum rotation angle of the rotatable ceramic coated sta...

Page 111: ...tation clamp screw and then rotate the stage by 180 3 If the target moves away from the center of the field of view turn the two stage centering screws with a hex driver to move the target halfway back towards the center 4 Use the stage knobs to move the target back to the center of the field of view 5 Bring the 40x objective into the optical path and repeat steps 2 to 4 The two stage centering sc...

Page 112: ...l Horizontal movement limit position A limit position of X direction and Y direction movement of the motorized XY stage is set as shown below by factory default X axis max value 34000 000 μm X axis min value 34000 000 μm Y axis max value 27000 000 μm Y axis min value 27000 000 μm The limit position can be changed programmatically See 23 Operation on DS L3 23 1 Setting Up the Microscope 6 Configuri...

Page 113: ...Procedure to focus on the specimen using the 10x objective 3 Operate the nosepiece to bring the 40x objective into the optical path and turn the coarse focus knob and then the fine focus knob to focus on the sample 4 Bring the 10x or 4x objective into the optical path 5 Look into the right eyepiece with your right eye without operating the focus knob and then focus on the sample by turning the rig...

Page 114: ...enter of the field diaphragm image is not centered turn the condenser centering screws to move the field diaphragm image to the center of the field of view This is easiest if you adjust the field diaphragm aperture so that it is slightly smaller than the eyepiece field of view Oil type condenser When using the D CUO DIC Condenser Oil apply immersion oil between the slide glass and the tip of the c...

Page 115: ...reases resolution and brightness but reduces contrast and depth of focus These characteristics involve inherent tradeoffs and cannot be optimized independently Proper size of the aperture diaphragm Calculating the aperture diaphragm diameter Use the following formula to determine the suitable size of the aperture diaphragm 34xNAx ratio NA Numerical aperture of objective Indicated on the side of th...

Page 116: ...perture diaphragm is fully opened Adjust the aperture diaphragm lever so that the size of the aperture diaphragm becomes 70 to 80 of the pupil 40x 0 75 Plan 40x Nikon JAPAN WD Plan 40X 40x 0 75 WD Indication for 40x magnification numerical aperture 0 75 Proper numerical aperture 0 75 x 0 7 to 0 8 0 525 to 0 6 Adjusting the aperture diaphragm using the centering telescope 1 Remove one eyepiece and ...

Page 117: ...nly the bright field microscopy is available for 2x and 4x with a 2 4x auxiliary lens in the optical path when used DIC Condenser Oil C C Achromat Swing out Condenser 1 100x 2 2 2 2 Swing out the top lens before use C C Slide Achro Condenser 2 100x 3 3 3 Do not use this condenser with an achromat objective or a plan achromat objective 3 For objectives of 10x or higher magnification pull out the sl...

Page 118: ...ed on the PH objective Dark field module required for dark field microscopy The dark field module D C DF can be used with any objective with NA numerical aperture of 0 7 or less Note however that 2x and 4x objectives are not supported 2 4x auxiliary lens required for low magnification bright field microscopy 2x and 4x objectives for universal condenser can only be used for bright field microscopy ...

Page 119: ...t Microscopy Procedure 14 Center the PH module Motorized universal condenser viewed from left rear side Operation from DS L3 See 23 2 Controlling the Microscope 9 3 Using the NI CUD Universal Condenser Dry The NI CUD is a dry type manual universal condenser that supports various microscopy methods Optical module The turret of the universal condenser has one empty hole and six holes for attaching o...

Page 120: ...nges the size of the aperture diaphragm Turn the diaphragm clockwise to open and counterclockwise to close A scale indicating the numerical aperture of the aperture diaphragm of the condenser is displayed at the top of the aperture diaphragm lever Match the indicator of the aperture diaphragm lever to the scale See 8 Adjusting the Aperture Diaphragm Aperture diaphragm at phase contrast microscopy ...

Page 121: ...ith the DIC module attached in the condenser slot during differential interference contrast microscopy Select the DIC module appropriate for the microscopy method and objective If the combination of the objective and DIC module is not correct differential interference contrast image cannot be obtained or the contrast decreases significantly Depending on the purpose of observation dedicated modules...

Page 122: ..._0 000um BF____AS30 6mm__FS30 0mm Right size of the field diaphragm Proper size of the field diaphragm Usually the size is optimal when it circumscribes the field of view Opening the field diaphragm too much and illuminating a broader area than necessary will result in stray light entering the field of view generating flare and reducing the image contrast In addition the specimen will become decol...

Page 123: ...a specimen from contacting due to the rotation of the motorized nosepiece Z position nosepiece rotation stop Since the focal depth of the objective with low magnification is deep the objective may get too closer to a specimen during microscopy When the objective is switched to a high magnification objective in this case the tip of the objective may contact the specimen This is to prevent the dange...

Page 124: ...stop is disabled by factory default Perform configuration on DS L3 See 23 Operation on DS L3 23 1 Setting Up the Microscope 4 4 Nosepiece Reverse Rotation for details Toggling the motorized nosepiece A sample can be observed by microscope while switching two objectives set in advance alternately using the toggle function Use the OBJ CW CCW button to switch the objectives You can set two patterns f...

Page 125: ...F parfocal correction See 19 2 Interlocking Function for conditions for interlocking and interlocked operations 11 2 Intelligent Manual Nosepiece Operation To switch objectives using an intelligent nosepiece or manual nosepiece manually turn the nosepiece Turn the nosepiece correctly so that it comes to the click stop position Attach the objective so that the magnification increases when the nosep...

Page 126: ...t 100 to NI RPZ NI RPZ EDSC DSC zooming port Motorized optical path switching PA__2x____Z _____0 000um BINO____ZM 1 0x_FS30 0mm Operation on DS L3 See 23 2 Controlling the Microscope Interlocked operation of optical path switching The optical path switching of the motorized quadrocular tilting tube can be interlocked when pressing the CAPTURE button to capture an image See 19 2 Interlocking Functi...

Page 127: ...lled out 0 100 Ridding the click feeling of the optical path switching C TT Trinocular Tube T and C TF Trinocular Tube F have a NO CLICK switch on their tube attaching surface Slide this switch in the direction of the arrow with the tip of a pointed tool to disable clicking for the optical path switching lever Set the switch to this position if you need to eliminate the slight vibrations resulting...

Page 128: ... section can be extended along the optical axis Adjust the position of the binocular section for most comfortable viewing Adjustment range Ergonomic tube 10 to 30 Quadrocular tilting tube 15 to 35 Extension range Ergonomic tube 40 mm 50 60 70 Adjusting the binocular section 13 2 Using the Eyelevel Riser Inserting the eyelevel riser between the tube and the microscope arm allows you to bring the ey...

Page 129: ...immersion oil Be careful not to apply immersion oil between other dry condensers and a specimen when used Air bubbles in the oil Any bubbles in the immersion oil will degrade image quality Be careful to prevent bubbles from forming To check for air bubbles fully open the field diaphragm and aperture diaphragm remove the eyepiece and examine the exit pupil bright round section of the objective insi...

Page 130: ... the water since excess water will flow onto the stage and around the condenser promoting corrosion In such cases immediately wipe off the water Cleaning of water After use wipe off the water from the tip of the objective and condenser then follow up by wiping with absolute alcohol If you observe water stains apply a small amount of neutral detergent and wipe gently and then follow up with absolut...

Page 131: ...ary specimen Oil immersion of a condenser When using a dark field condenser oil apply immersion oil between the slide glass and the condenser lens See 14 Oil Water Immersion 5 Turn the condenser focus knob slowly to lower the condenser lens until a dark shadow appears near the center of the field of view 6 Using the condenser centering screws move the dark shadow to the center of the field of view...

Page 132: ...rescence cube turret operation Use a FL CUBE CW CCW button to switch the epi fluorescence cube turret The turret rotates to the adjacent address each time the button is pressed Attachment information display for the filter cube is disabled by factory default Set the filter cube attachment information to the epi fluorescence cube turret and switch it The information on the filter cube on the optica...

Page 133: ...scence cube turret The switching of the motorized epi fluorescence cube turret can be interlocked when pressing the CAPTURE button to capture an image See 19 2 Interlocking Function for conditions for interlocking and interlocked operations Intelligent epi fluorescence cube turret epi fluorescence cube turret operation Manually rotate the cube switchover turret to switch the filter cube between in...

Page 134: ... wavelengths allowed to pass through a filter is referred to as the bandwidth The bandwidth range of an excitation filter determines the brightness of the fluorescent image the generation of auto fluorescence fluorescence resulting from substances other than the fluorophores and degree of fading The broader the bandwidth the greater the amount of excitation light irradiated onto the specimen there...

Page 135: ...mance have resulted in increased use of filters of short cut on wavelengths 2 For multiple labeled specimens use an LP filter for microscopy of fluorescent images of all fluorophores Note that a combination involving an ordinary dichroic mirror an excitation filter and an LP filter type barrier filter will be incapable of excited fluorophores that emit long wavelength fluorescent light for example...

Page 136: ... shutter Opening and closing the shutter of a motorized unit ND 8 ND 32 OUT IN NCB 11 Opening closing the shutter BA400_____DAPI_ 2 C_____ EX330 380_UV 2B C__EPI C PA__2x____Z _____0 000um UV 2B C____ND_1 C__EPI C C shutter closed O shutter open Motorized epi fluorescence cube turret Press the FL SHUTTER button to open close the shutter of a motorized epi fluorescence cube turret The shutter is cl...

Page 137: ...and the DIA motorized shutter on the dia illumination side to the DIA SHUTTER connector These are set by factory default Change the setting if you intend to use the motorized shutter for AUX other than EPI DIA Change the configuration on DS L3 See 23 Operation on DS L3 23 1 Setting Up the Microscope 2 Configuring the Device Connection Information for details Motorized HG precentered fiber illumina...

Page 138: ...intensity The higher filter number the lower transmittance and darker images are produced when placed in the optical path ND filters do not affect the color balance Image brightness can also be adjusted by using the aperture diaphragm on the epi fluorescence attachment ND filters in the epi fluorescence attachment The epi fluorescence attachment has three ND filters ND4 ND8 and ND16 built in If th...

Page 139: ...Under epi fluorescence microscopy it is also used to adjust the brightness of the image as well as the size of stray light Turning the aperture diaphragm lever of the epi fluorescence attachment changes the size of the aperture diaphragm Pushing in the lever opens the aperture diaphragm while pulling it out closes it Stopping down the aperture diaphragm reduces stray light but will also make the i...

Page 140: ...escence attachment as follows before use Center the field diaphragm 1 Pull out the field diaphragm lever on the epi fluorescence attachment to stop down the field diaphragm 2 Turn the field diaphragm centering screw with the provided hex driver 2 mm across flats to move the field diaphragm image to the center of the field of view 3 Push in the field diaphragm lever and open the field diaphragm to ...

Page 141: ...he glass surface of the excitation balancer 4 Attach the aperture diaphragm unit back onto the epi fluorescence attachment 5 Attach a dual band filter cube onto the epi fluorescence attachment 6 Adjust the excitation balancer horizontally to attain the desired excitation light Attaching the excitation balancer Excitation balancer details The excitation balancer is configured to achieve almost 100 ...

Page 142: ...h excitation filter See Chapter 1 3 Assembly Method 13 Attach the motorized excitation filter wheel in the Assembly Maintenance instruction manual for attaching the excitation filter Configure the information on attaching the excitation filter to the wheel on DS L3 See 23 Operation on DS L3 23 1 Setting Up the Microscope 1 Setting Optical Elements Installation Information for details The wheel is ...

Page 143: ...l Filter nameplates can also be attached onto the replacement cover for your convenience Operation on DS L3 See 23 2 Controlling the Microscope 5 A λ 5 4 3 2 1 ND4 ND8 ND16 1 1 O 2 3 4 5 6 7 8 50 60 70 Switching of barrier filter BA400_____DAPI_ 2 C_____ EX330 380_UV 2B C__EPI C 16 9 Other Notes on Epi fluorescence Microscopy Using a non fluorescent slide cover glass and immersion oil For fluoresc...

Page 144: ... this adjustment Adjust the orientation of the analyzer attached to the nosepiece and the polarizer on the field lens The orientation of the analyzer is fixed in Ni E Adjust the orientation of the polarizer to find a position where field of view is darkest See Chapter 2 5 Differential Interference Contrast Microscopy Procedure 14 Adjust the orientation vibration direction of the polarizer and anal...

Page 145: ...click positions The slider will click once after insertion into the slot At this point the lambda plate is not in the optical path To bring the analyzer into the optical path push the slider further to the second click stop position This will change the background to a sensitive color enabling observation with high color contrast An interference color is displayed based on changes in the refractio...

Page 146: ... below for the use properties of Ph objectives When using a dark contrast Ph objective make sure that the phase contrast of the specimen does not exceed the allowed phase contrast amount latitude If the phase contrast amount of the specimen is greater than the allowed phase contrast amount observation is not possible as the image will be illuminated brighter than the background You can increase or...

Page 147: ...hase plate they may present images that differ somewhat from those presented by method specific objectives Use method specific objectives for strict observations Centering of the objective phase plate and the PH module Ph annular diaphragm A phase contrast effect is achieved by correctly aligning the objective s phase plate with the condenser s PH module Ph annular diaphragm The phase image will d...

Page 148: ...l Up to four alphanumeric characters can be used for a mode name Here MODE1 is named as mdic and MODE2 as mfl for registration 1 Registering a mode Start from the DS L3 MICROSCOPE SETUP screen SETUP MENU MODE 1 Specify MODE1 For the initial registration of the mode factory setting mode names MD1 to MD8 are displayed on the buttons The mode name is displayed on the button for which a mode has been ...

Page 149: ...n the right of the registered MODE mdic on a MODE registration complete screen The current state of the registered device will be saved 3 Perform the epi fluorescence microscopy to get ready for save 4 Press the SAVE MODE2 button for MODE mfl The current state of the registered device will be saved Saving a mode Operation on the MICROSCOPE CONTROL screen 1 Perform the differential interference con...

Page 150: ...ION button function You can work with on the DS L3 MICROSCOPE CONTROL screen Press the LOAD MODE button Press mdic and mfl alternately when a sub screen of the mode selection load opened Saved differential interference contrast microscopy and epi fluorescence microscopy will be loaded alternately MICROSCOPE CONTROL Loading MODE Loading a mode in Sleep mode Loading a mode is not available in Sleep ...

Page 151: ...er of the motorized barrier filter wheel in the optical path PATH Optical path in the motorized quadrocular tilting tube Z Position of the elevating section 1 EPI SHUTTER Opening closing of the EPI motorized shutter DIA SHUTTER Opening closing of the DIA motorized shutter LAMP ADJ Brightness of the dia illumination lamp LAMP Turns the dia illumination lamp on or off INTSL SHUTTER Opening closing o...

Page 152: ...NT 2 OFF factory setting Requires to connect a motorized intelligent nosepiece that requires defined objective information motorized quadrocular tilting tube motorized ND filter wheel and a motorized universal condenser and the aperture diaphragm should be open 75 If Intelligent OPEN or CLOSE is set for the aperture diaphragm the transmittance of ND filters to be adjusted may not be optimized In t...

Page 153: ...e A STOP F STOP and ND will be reset once DS L3 is powered off In order to use the INTELLIGENT setting for the next microscopy be sure to press the SAVE button to save the settings Restore the changed default of INTELLIGENT to the factory setting When the information of the object is reset the default INTELLIGENT of that nosepiece address is restored to the factory setting See Chapter 3 23 Operati...

Page 154: ...CTIVE INTERLOCK this is to set either Interlock Do not interlock for those movement involved in zooming It is set to Do not interlock as factory default Interlocking in Sleep mode No interlocking takes place in Sleep mode Interlocking with optical path switching When the field diaphragm has been set in the OBJECTIVE INTERLOCK this is to set either Interlock Do not interlock for the movement of the...

Page 155: ...extra fine 2 XY speed switch LED Switches the operation mode of the stage knobs between coarse fine and extra fine LED off coarse LED on fine LED blinking extra fine 3 CW button clockwise Controls clockwise rotations The control target is selected with 8 4 CCW button counterclockwise Controls counterclockwise rotations The control target is selected with 8 5 FL SHUTTER button LED Opens closes the ...

Page 156: ... BINO 100 to binocular Rotate epi fluorescence cube turret on the 2nd layer Switch optical path FRONT 100 to tube adapter Rotate nosepiece Switch optical path REAR 100 to rear port Switch ND filter value in the HG precentered fiber illuminator Switch optical path B100 F100 Rotate condenser turret Switch optical path F100 R100 Adjust aperture diaphragm on the condenser Switch optical path R100 B100...

Page 157: ...te a motorized XY stage as if it were a manually operated stage Pressing the XY Speed switch on the operation panel of the ergo controller will switch the operation mode of the knobs between coarse fine and extra fine with coarse being the factory setting The operation mode is indicated by the switch LED off coarse on fine blinking extra fine Orientation of the stage knobs The relative position of...

Page 158: ...mode will be indicated by the Coarse Fine or ExFine XY Speed LED XY Speed LED Coarse Fine or ExFine will light up to indicate the currently selected stage XY movement mode Constant Speed switch Sets the stage XY movement to constant speed Pressing this switch while operating the stage with the joystick will fix the stage movement speed at the speed at that point Under constant speed operation the ...

Page 159: ... image with a camera cable connected to DS L3 controlled with the USB CAPTURE button commands the image capture The CAPTURE button function is set to capture an image using a camera connected to the USB as default Operation on DS L3 See 23 2 Controlling the Microscope IN ND 8 ND 32 NCB 11 OUT 50 60 70 Interlocking setting caused by CAPTURE Interlock Do not interlock setting is available for the op...

Page 160: ...or the C mount so that an image moves in the reverse direction when the stage is moved right and left adjust the camera position and then tighten the screw to fix it 3 Focus the image If the image viewed through the eyepiece appears to be in focus but the image on the monitor is out of focus make adjustment Adjusting the camera position for the DSC zooming port CONTACT ARM2 Focusing the image usin...

Page 161: ...rovided with the camera Number of images captured Serigraphy is not available because the CAPTURE button is designed to capture only one image ZOOM Attaching a camera to the zooming port allows to zoom in an image Motorized DSC Zooming Port The final magnification of the motorized DSC zooming port can be from 0 6x up to 2 0x Use the CW CCW button of the ergo controller for a zoom operation A zoom ...

Page 162: ...ck feeling of the zoom operation Bottom of the zooming port Limited zoom range Note that the zoom range may be limited depending on capture conditions or type of the camera when using a DS camera made by other manufacturers Limb darkening in zooming port When performing bright field microscopy with a tube directly attached to the arm the edge of the field of view may be dark at zoom magnification ...

Page 163: ...may fade during exposure To prevent this do the following Use a brighter optical system combination Even if the overall magnification is the same on the monitor the combination of objective and camera zoom can result in significant variations in exposure time Nikon recommends increasing the magnification of the objective rather than of the zoom Generally the numerical aperture of the objective inc...

Page 164: ...ning ON DS L3 displays the top screen of the camera control at first Use the button to navigate to the MICROSCOPE CONTROL top screen Camera Control top screen MICROSCOPE CONTROL Top Screen Using M SETUP button to navigate to MICROSCOPE SETUP MENU screen 2 Display the MICROSCOPE SETUP MENU screen MICROSCOPE SETUP MENU screen Selecting a Setup Menu 3 Select a setup menu Press your desired menu butto...

Page 165: ...e cube turret SETUP MENU COMPONENTS Device selection Device icons on the top of the screen are as shown below from left Motorized nosepiece intelligent nosepiece Objective settings Motorized condenser DIC module and PH module settings Motorized intelligent epi fluorescence cube turret on the 1st layer Filter cube settings Motorized intelligent epi fluorescence cube turret on the 2nd layer Filter c...

Page 166: ...cable to capture image by sending trigger signal from the DSC connector on the main body This configuration is unnecessary if you are capturing from a camera directly connected to DS L3 by pressing the CAPTURE button on the camera control screen on DS L3 Default setting NOT CONNECTED 2 2 Connecting the motorized shutter See DS L3 Chapter 6 3 2 Configuring the Connection of Motorized Shutter Perfor...

Page 167: ...CTION button Function setting Factory setting Buttons 1 and 2 Rotates the condenser clockwise and counterclockwise Buttons 3 and 4 Rotates the excitation filter wheel clockwise and counterclockwise Buttons 5 and 6 Rotates the motorized barrier filter wheel clockwise and counterclockwise Press the FUNC SET button under FUNCTION BUTTON to specify the button number and to select a function Selecting ...

Page 168: ...USB Camera directly controlled by DS L3 connected to the USB connector default setting NOT CONNECTED FRONT LEFT AUX Camera connected to DSC1 at the position set in 2 Configuring the Device Connection Information CAPTURE Button NOT CONNECTED FRONT LEFT AUX Camera connected to DSC2 at the position set in 2 Configuring the Device Connection Information L FL R OBJ OBJ CW CCW button on the lateral left...

Page 169: ...er displays the button alignment of the ergo controller Specify a button to select a function The CW and CCW buttons in the middle have different functions available for setting than other buttons Selecting supported unit Ex When a upper left button is pressed 3 3 Buttons on the Screens on DS L3 See DS L3 Chapter 6 4 1 Configuring the Screen Buttons for DS L3 Configure the buttons on DS L3 screens...

Page 170: ...ture diaphragm built in the motorized universal condenser Motorized DSC zooming port Motorized ND filter wheel The following operation is not available in Sleep mode Loading MODE Grouped operation of EPI side shutters Showing hiding the sleep button The message Sleep mode is active Drive operation not available as shown in the figure on the right side is displayed on the Ni E front display panel a...

Page 171: ...6 5 1 1 Configuring the Interlocked Operation with Switching of Objectives See DS L3 Chapter 6 5 1 2 Changing the Initial Value of the INTELLIGENT See DS L3 Chapter 6 5 1 3 Automatically Switching the Movement Speed of the Microscope s Elevating Section and Motorized XY Stage Set a movement that interlocks with switching of objectives OBJECTIVE INTERLOCK setting Set the interlocking for each item ...

Page 172: ...with the Intelligent settings of the Aperture Diaphragm Field Diaphragm and ND Value will be reset once DS L3 is powered off In order to use the Intelligent setting for the next microscopy be sure to press the SAVE button to save the settings SAVE in OBJECTIVE INTERLOCK setting ZOOM INTERLOCK See DS L3 Chapter 6 5 1 5 Configuring the Interlocked Operation with Switching of Zoom Magnification When ...

Page 173: ... path or epi fluorescence cube turret to set the interlocking Specifying target to interlock with image capture Interlock setting Example epi fluorescence cube turret 4 2 Escape Distance See DS L3 Chapter 6 5 2 Setting the Retracting Amount of the Elevating Section Set how much the stage lowers from the current position when pressing the ESCAPE button of the microscope or the OBJECTIVE INTERLOCK h...

Page 174: ...Z POS NOSEPIECE ROTATION STOP is automatically reset once DS L3 is powered off even if the SAVE button is pressed on the MAIN screen to save the settings Effect of the vibration When vibration should slightly move the elevating section around the setting value the motorized nosepiece may rotate Specify Z Pos nosepiece rotation stop Button is checked when the rotation stop is set Pressing the butto...

Page 175: ...ached Factory setting NOT LIMITED 4 5 Nosepiece Toggle See DS L3 Chapter 6 5 5 Configuring Toggle Function Alternating between Two Objectives A sample can be observed by microscope while switching two objectives set in advance alternately using the toggle function Use the Rotate nosepiece Rotate nosepiece reversely button for the microscope main body or ergo controller to switch the objectives You...

Page 176: ...apter 5 2 2 Saving Updating a Mode State of Motorized Devices After the mode has been registered the mode name registered for the MODE No button is displayed and the SAVE MODE button is displayed on the right of the button Let the microscope enter your desired microscopy state here press the SAVE MODE button and then the current position and state mode of the registered motorized unit will be save...

Page 177: ...ttern by pressing the DISPLAY buttons on the Ni E main body then press the OK button Display switching between microscope state and FUNCTION button s function Pressing the DISPLAY switch button switches the status display and the function display of the FUNCTION button SETUP MENU UTILITY Display pattern setting 6 2 Enabling Z Control See DS L3 Chapter 6 6 2 Enabling Disabling the Operation of the ...

Page 178: ... Set upper lower limit position For focusing stage system the upper limit is set 0 5μm above the current position and for focusing nosepiece system the lower limit is set 0 5μm below the current position If the elevating section is currently at the limit position it cannot be returned to default factory setting In this case move the elevating section below the limit position above the limit positi...

Page 179: ...ck the program version of the microscope system When you are using an ergo controller the program version of the ergo controller is displayed after ERGC When you are using the joystick controller the program version of the joystick controller is displayed after JOY SETUP MENU MAINTENANCE Maintenance Item Setting Restoring the Factory Default Settings for Microscope Data from the Microscope You can...

Page 180: ...rret 3 Switching of filter cube opening closing of internal shutter NI FLT6 I intelligent epi fluorescence cube turret 3 Address detection for turret in optical path Information about the attached filter cube See Chapter 6 2 4 Configuring the Excitation Filter Barrier Filter Information NI EXW E motorized excitation filter wheel Switching of excitation filter NI BAW E motorized barrier filter whee...

Page 181: ...sepiece FL turret etc Press the button to open a sub screen at the bottom of the screen X X coordinate of the motorized XY stage Y Y coordinate of the motorized XY stage Z Z coordinate of the microscope s elevating section Closes the MICROSCOPE CONTROL screen Operation buttons optical path etc Press a button to operate the corresponding motorized device The selected button is displayed with a blue...

Page 182: ...s you to control the following parts of Ni E or motorized devices See the list for the function and descriptions on the buttons IN ND 8 ND 32 NCB 11 OUT 1 2 A λ F STOP A STOP EX ADJ 2 3 4 5 6 1 1 O 2 3 4 5 6 7 8 50 60 70 INTENSILIGHT C HGFIE LAMP POWER RUN TIME hrs EPI for epi illumination DIA for dia illumination ...

Page 183: ... turret 1st layer Chapter 5 1 4 Switching the Filter Cube Motorized Epi fluorescence Cube Turret FL TURRET SHUTTER FL Open close the motorized epi fluorescence cube turret s built in shutter 1st layer Chapter 5 1 5 Opening Closing the Motorized Epi fluorescence Cube Turret s Built in Shutter Filter Cube 2nd Address Switch the filter cube of the motorized epi fluorescence cube turret 2nd layer Chap...

Page 184: ...just the brightness of the dia illumination lamp PHOTO Adjust the dia illumination lamp to the brightness that offers optimal color reproduction Chapter 5 1 14 3 Adjusting the Brightness of the Dia illumination Lamp LED LAMP LAMP ON OFF Turn the dia illumination lamp ON OFF Chapter 5 1 14 2 Turning the Dia illumination Lamp LED ON OFF F STOP Adjust the diameter of the DIA field diaphragm Chapter 5...

Page 185: ...utput a capture trigger signal to a digital camera connected to the position other than the above Chapter 5 1 17 Outputting Capture Trigger Signals from the Microscope SAVE MODE Save the current microscopy state as a mode Chapter 5 2 2 Saving updating a Mode State of Motorized Devices LOAD MODE LOAD MODE number Load a saved mode Chapter 5 2 3 Loading a Mode Z axis RESET Reset the Z axis coordinate...

Page 186: ...Operation Reference in DS L3 Instruction Manual SLEEP Enter the sleep state to reduce noise Chapter 5 3 Entering the Sleep State Noise Reduction COLOR Change the background color of the MICROSCOPE CONTROL screen Chapter 5 4 Changing the Background Color of the MICROSCOPE CONTROL Screen ...

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