Motic AE30-31 Instruction Manual Download | Manualshive

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Motic Instruments Inc. Canada

October 06, 2006

37

Motic

Standard Fluorescence Filter Sets

MF31000

DAPI and Hoechst Set:
Exciter D350/50x
Dichroic 400DCLP
Emitter D460/50m

MF31001

FITC FITC/RSGFP/Fluo 3/DiO Acradine RNA) set:
Exciter D480/30x
Dichroic 505DCLP
Emitter D535/40m

MF31002

TRITC (Rhodamine)/DiI/Cy3 Set:
Exciter D540/25x
Dichroic 565DCLP
Emitter D605/55m

MF31004

Texas Red® / Cy3.5 Set:
Exciter D560/40x
Dichroic 595DCLP
Emitter D630/60m

MF41008

Cy5, Alexa Fluor 633, Alexa Fluor 647 Set:
Exciter HQ620/60x
Dichroic Q660LP
Emitter HQ700/75m

The emission at these wavelengths will be better detected by camera than by the
unaided eye. Only a small percentage of humans can detect these wavelengths.

MF31044

Cyan GFP Set:
Exciter D436/20x
Dichroic 455DCLP
Emitter D480/40m

This set will typically exclude the signal from YFP but not from GFP.

MF41017

Endow GFP Bandpass Emission Set:
Exciter HQ470/40x
Dichroic Q495LP
Emitter HQ525/50m

This is the recommended filter set for the newer S65T GFP mutants. It is also
recommended for wtGFP for which it was originally designed.

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Summary of Contents for AE30-31

Page 1: ...MOTIC INCORPORATION LTD Prepared by Motic Instruments Inc Canada Episcopic Fluorescence Attachment EF INV ll Inverted Microscope AE30 31 Instruction Manual...

Page 2: ...Attachment used in conjunction with the Motic AE30 31 Inverted Microscope We are constantly endeavouring to improve our instruments and to adapt them to the requirements of modern research techniques...

Page 3: ...ht microscopy The radiation emitted by the light source first passes through the exciting filter and is incident to a short pass filter dichroic beam splitter The radiation is then reflected into the...

Page 4: ...otic Instruments Inc Canada October 06 2006 4 Table of Contents 1 0 Names of Component Parts 2 0 Setting up the Instrument 3 0 Assembly 4 0 Microscopy 5 0 Troubleshooting Table 6 0 Care and Maintenanc...

Page 5: ...otic Instruments Inc Canada October 06 2006 5 1 0 Names of Component Parts Fig 1 Epi fluorescence attachment EF INV ll Light source Epi fluorescence attachment Light protective shield Power supply uni...

Page 6: ...n the microscope manual For the purpose of performing simultaneous observation with phase contrast attach the appropriate phase contrast components referring to the specific instructions provided 3 2...

Page 7: ...st covers See Figure 2 Pull out rear dust cover located above the Microscope AC inlet Fig 2 See Figures 3a and 3b Remove side dust covers located on each side of the microscope base Fig 3a Fig 3b Micr...

Page 8: ...See Figure 4 Attach the main body sleeve of the epi fluorescence attachment into the microscope stand with the notch on the sleeve facing left Fix the main body sleeve in this position by using the th...

Page 9: ...n the pin on the Epi fluorescence attachment with the notch in the Epi fluorescence attachment sleeve slide the main body of the Epi fluorescence horizontally into the sleeve until it cannot be pushed...

Page 10: ...slider into the opening of the main body of the fluorescence attachment Tighten the loosened knob Fig 6 Light cut off slider To prevent the specimen from fading during temporary pauses in observation...

Page 11: ...e 7 Slide the filter cassette into the opening on the right side of the microscope until the two pins slot into the two matching holes on the microscope opening surround Using an Allen hex key secure...

Page 12: ...th the opening on the left side of the microscope Slot the two pins into the two matching holes on the microscope opening surround Using an Allen hex key secure the filter cube cover retaining plate f...

Page 13: ...slot empty as strong light may enter and damage the user s eyes when shifting between excitation methods If fewer than three filter cubes are required for microscopy Use a DIA ILL dummy cube to ensure...

Page 14: ...filter slider Undo the Allen hex screws holding the clamp plate on right side of the slider with the hexagonal key supplied with the fluorescence attachment and secured inside the right filter cube c...

Page 15: ...nner Remember the position of the filter cube so that you can insert the ID tag supplied with the filter cube for the cube added Install frequently used filter cubes at the end of the dovetail mount i...

Page 16: ...itation method changeover lever into the filter cube through the opening in the white nylon bushing Fig 12b on the right cover Fig 13 Insert the filter cube ID tags Fasten the excitation method change...

Page 17: ...not in use store in a desiccator or a sealed container with a drying agent as interference filter characteristics change if the filter is exposed to high humidity Do not allow dust to get onto the fi...

Page 18: ...he rubber cap that covers the collector lens focus knob Loosen the set screw at the base of the focusing knob with a 2 5mm Allen hex key While pulling out the collector lens focus knob insert the coll...

Page 19: ...s knob pin in focussing groove of the collector lens Tighten the set screw at the base of the focusing knob with a 2 5mm Allen hex key Tighten the cheese head slotted screw following lamp alignment an...

Page 20: ...t ring on the Epi fluorescence attachment Fig 16a Fig 16b Secure the bayonet ring by turning it as far as possible in the direction of arrow Bayonet ring Align the positioning pin on the lamp house fl...

Page 21: ...clamp screws at both ends of the lamp socket and remove dummy lamp installed Before handling the lamp read the handbook supplied with the lamp When installing the lamp avoid applying force that might...

Page 22: ...ver over the inner metal surround of the lamp house and pivot the lamp socket cover to close Secure with the clamp screw The safety cut off micro switch will click as you secure the lamp house cover c...

Page 23: ...sed in your area and that has been approved to meet local safety standards Using the wrong power cord could cause fire or equipment damage When using the extension cord only use a power supply cord wi...

Page 24: ...the power is turned on The POWER LAMP READY LAMP indicator lamp will start flashing briefly to indicate that the lamp is stabilized Press the RESET PAD below the RUN TIME counter on the power supply T...

Page 25: ...n turn ionize the gas in the bulb igniting the lamp These ions under low voltage carry the current that generates the light between the electrodes The lamp gets very hot during the vaporization of the...

Page 26: ...of the centering tool To begin alignment of the lamp rotate the collector lens focus knob to produce a sharp image of the arc on the window of the centering tool Rotate the lamp horizontal and vertic...

Page 27: ...tly outside the edge of the field of view If a larger than required area is illuminated extraneous light will enter the field of view This will create a flare in the image and lower the contrast Cutti...

Page 28: ...en the field aperture diaphragm just slightly outside the edge of the field of view Lower the field aperture diaphragm lever Fig 22 3 20 Adjusting the shutter slider Whenever observation is interrupte...

Page 29: ...ter Blocks A cube shaped modules that hold a matched set of fluorescence filters including excitation EX barrier emission BA and a dichroic mirror DM The excitation filter allows transmission of light...

Page 30: ...so leads to a high level of self fluorescence and severe fading To minimize spectral overlap one can instead choose excitation and emission filters that are narrow in bandwidth and are spectrally well...

Page 31: ...Filters that pass light of a certain restricted range of wavelength Bandpass filters are useful for observing a certain stain on multi stained specimen such as when two stains FITC and TRITC are used...

Page 32: ...oil container before use Immersion oil must be used sparingly Freedom from air bubbles must be ensured To check for air bubbles remove one eyepiece open field diaphragm as far as possible and look at...

Page 33: ...tter is inversely proportional to the magnification To minimise the magnification required to fill the picture with the desired object the format chosen should be as small as practicable and hence hav...

Page 34: ...ce life Lamp not securely plugged into socket Optical Image not visible Shutter slider in light path Filter cube unsuitable for specimen Filter cube not completely in light path Lamp on but image is u...

Page 35: ...rs deteriorate with time due to exposure to heat and intense light Replace when necessary Handle Multi Band filters with great care as the very complex coatings wear easily Special attention should be...

Page 36: ...dirt moisten a piece of gauze with diluted detergent and wipe clean 6 3 When not in use When not in use cover the instrument with vinyl dust cover and store in a place low in humidity where mold is un...

Page 37: ...D630 60m MF41008 Cy5 Alexa Fluor 633 Alexa Fluor 647 Set Exciter HQ620 60x Dichroic Q660LP Emitter HQ700 75m The emission at these wavelengths will be better detected by camera than by the unaided ey...

Page 38: ...41028 Yellow GFP BP 10C Topaz Set Exciter HQ500 20x Dichroic Q515LP Emitter HQ535 30m This set is designed to image YFP It will not in most cases detect CFP when co expressed in cells By eye fluoresce...

Page 39: ...uments Inc Canada October 06 2006 39 Arrangement of Filters in a Fluorescence Cube Inverted Microscope Dichroic Mirror Lamp HBO 100W Exciter Filter Barrier Filter To Observation Method To Objective an...

Page 40: ...r is reflected 90 toward the objective and the specimen Finally light emanating from the specimen is passed through and directed toward the emission filter and observation method Dichroic multi layer...

Page 41: ...FWHM Example D350 50x is an interference filter with maximal transmission at 350nm and transmits light from 325 and 375nm See Figure Below GFP The Green Fluorescent Protein GFP from the jellyfish Aequ...

Page 42: ...rs reflect and transmit more than two bands of light Dual band or Triple band filter sets includes three carefully balanced combinations that contain dual or triple bandpass excitation and emission fi...

Page 43: ...hortpass and longpass filters block or transmit wavelengths at specific cut off wavelengths Bandpass filters exhibit broadband or shortband transmission centered on a particular band of wavelengths Fi...

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