background image

22

23

Brightness (%)

Wavelength (nm)

For fluorochrome emission, a light beam having a specific wavelength is selected from a 
wide spectrum of wavelengths. The five major peaks of luminance are at wavelengths of 
365/366, 404.7, 435, 546.1 and 577.0/579.1 nm. In addition, light beams having wavelengths 
of  334.2  and  490  nm  (at  rather  low  luminance)  are  also  applicable  to  fluorochrome 
emission.

Typical Example of Emission Spectrum of Ultrahigh-Vacuum

Mercury Burner Spectrum

17.  For red fluorescent protein (RFP)

U-FRFP

Tr

ansmittance (%)

Wavelength (nm)

18.  For mCherry

U-FMCHE

Tr

ansmittance (%)

Wavelength (nm)

Содержание BX3-RFAS

Страница 1: ...rize yourself fully with the use of this system we recommend that you study this manual thoroughly before operating the microscope Retain this instruction manual in an easily accessible place near the work desk for future reference A X 7 8 8 8 BX3 URA BX3 RFAS U LH100HGAPO U LH100HG U RFL T U AN 2 BX3 25ND6 BX3 25ND25 REFLECTED FLUORESCENCE SYSTEM ...

Страница 2: ... complies with the requirements of both directive 2004 108 EC concerning electromagnetic compatibility and directive 2006 95 EC concerning low voltage The CE marking indicates compliance with the above directives ...

Страница 3: ...RFAS BX3 URA 1 General Precautions for Observation 9 2 Selecting the Fluorescence Mirror Unit 9 3 Turning the Power Supply Unit On 11 4 Centering the Field Iris Diaphragm 11 5 Centering the Aperture Iris Diaphragm 12 6 Centering the Mercury Burner 13 1 Simultaneous Reflected Fluorescence and Phase Contrast Observations 18 8 1 Assembly Diagram 24 8 2 Detailed Assembly Procedures 25 2 7 Mounting the...

Страница 4: ...ousing for replacement of the burner or any other internal part set the main switch to OFF then unplug the lamp housing connection cable from the power supply unit and wait for more than 10 minutes until the lamp housing cools down This system employs a UIS2 Universal Infinity System 2 optical design and should be used only with UIS2 microscopes eyepieces objectives and condensers for the BX3 seri...

Страница 5: ...eflected fluorescence system Before using this system together with the BX3 microscope and associated options make sure that you have carefully read and understood their manuals and understand how the system should be operated together 2 The reflected fluorescence system is composed of precision instruments Handle it with care and avoid subjecting it to sudden or severe impact 3 Do not use the sys...

Страница 6: ...rs USH 103OL HBO103W 2 When the hour counter on the power supply unit indicates this value set the main switch to OFF and wait for more than 10 minutes before replacing the mercury burner Page 26 Unlike electric bulbs the mercury burner seals high pressure gas inside If it continues to be used after the service life has expired the glass tube may eventually explode due to accumulated distortion 5 ...

Страница 7: ...sides Page 11 Shutter OPEN Shutter CLOSE Field iris diaphragm knob Page 11 Analyzer insertion slot Page 18 Field iris diaphragm centering screws Page 11 x2 screws Collector lens focusing knob Page 13 14 Burner centering knobs Page 13 14 Mirror focusing screw Page 15 On the rear of the lamp housing ND filter insertion slots Page 16 Provided on both sides Aperture iris diaphragm centering screws Pag...

Страница 8: ...Shutter OPEN Shutter CLOSE ND filter slider insertion slot Page 16 Insertion from right side Aperture iris diaphragm knob Page 12 Field iris diaphragm knob Page 11 Field iris diaphragm centering screws Page 11 Analyzer inlet Stopper ND filter slider for BX3 RFAS Engaged filter name inscription Stopper Collector lens focusing knob Burner centering knobs Mirror focusing screw On the rear of the lamp...

Страница 9: ...indicator sheets provided with the illuminator can be used to write the names of original fluorescence mirror units Power Supply Unit U RFL T U RX T ND Filters BX3 25ND6 BX3 25ND25 Centering Target U CST For details see the instruction manual provided with the U RFL T U RX T Use the U RFL T when the light source is a mercury burner or use the U RX T when the light source is a xenon burner Hour cou...

Страница 10: ...h the phase contrast observation or transmitted light Nomarski Differential Interference Contrast DIC observation please read Chapter 4 SIMULTANEOUS FLUORESCENCE OBSERVATION Page 18 Controls Used Page Attach the fluorescence mirror unit and objective matching the observation method Pages 9 to 10 Center the mercury burner Pages 13 to 15 Adjust the field iris diaphragm Adjust the aperture iris diaph...

Страница 11: ...8 Make a photocopy of the observation procedure pages and post it near your microscope 9 a 7 5 2 8 3 6 b 1 4 ...

Страница 12: ...g of specimens This system features high excitation light intensity to ensure bright observation of dark fluorescence specimens In consequence after long period of observations using high power objectives the colors of specimens will fade quicker than usual causing the view contrast of fluorescent images to deteriorate In such a case slightly reduce the excitation light intensity to slow color fad...

Страница 13: ...P530 550 BA575IF Rhodamine TRITC Florescent antibody Propidium iodide DNA RFP Y U FYW DM600 BP545 580 BA610IF Texas Red Fluorescent antibody Combinations for color separation U U FUNA DM410 BP360 370 BA420 460 For observing only the U excitation stain when using U excitation stain together with FITC B U FBWA DM505 BP460 495 BA510 550 For observing only the B excitation stain when using B excitatio...

Страница 14: ... after use After replacing the mercury burner reset the hour counter by holding its reset button till 0 0 is displayed CAUTION CAUTION Fig 1 4 Centering the Field Iris Diaphragm Fig 1 1 Close the light path by sliding the shutter knob to position marked 2 Engage a mirror unit other than the ones for U excitation U FUW U FUN U FUNA in the light path by rotating the turret 3 Open the light path by s...

Страница 15: ...e center of field 6 Turn the revolving nosepiece to engage the empty place the objective cap should be removed in the light path 7 Pull out the aperture iris diaphragm knob 2 to minimize the aperture iris diameter 8 Pull out the field iris diaphragm knob 3 to minimize the field iris diaphragm Now the aperture iris image should be visible on the U CST 9 Fit the Allen screwdriver in the two aperture...

Страница 16: ...on the cross lines for reflected light white surface 5 While looking into the eyepieces adjust the stage to bring the center of the cross lines for reflected light on the white surface on the center of the field of view 6 Turn the revolving nosepiece to engage the empty position the objective cap should be removed in the light path 7 Open the shutter by sliding the shutter knob to position marked ...

Страница 17: ...the shutter by setting the shutter knob to position marked 8 While looking into the eyepieces turn the collector lens focusing knob to project the arc image and mirror arc image on the U CST A in Fig 7 The mirror arc image refers to the arc image projected from the reflection of the lighting section between the poles on the mirror built into the lamp housing If the poles are displaced excessively ...

Страница 18: ...nt has been executed the mirror can never be returned to the same status as the factory shipment status 1 Using a pair of tweezers etc peel off the two blind seals 6 from the rear of the lamp housing 2 Loosen the screws below the seals using the Allen screwdriver The mirror is unclamped when these two screws are loosened 3 Then peel off another couple of blind seals This exposes the mirror arc ima...

Страница 19: ...ted back to front with the filter name inscription facing the opposite direction to the observer the filter may be damaged The first click position of the filter engages the empty position in the light path and inserting the filter till the second click position engages the filter CAUTION Fig 9 Fig 10 1 3 2 1 3 2 4 Operation with BX3 RFAS Fig 10 Use the ND filter slider provided with the BX3 RFAS ...

Страница 20: ...nser and the area around it In this case use the light shield sheet 1 Lower the condenser by turning the condenser knob 2 Insert the light shield sheet into the space on the stage When switching the observation method to transmitted light observation phase contrast Nomarski etc the light shield sheet should simply be placed on the window lens 2 only during fluorescence observation 1 2 ...

Страница 21: ...al provided with the U UCD8 transmitted light universal condenser The transmitted light DIC observation requires the following accessories universal condenser U UCD8 2 transmitted light DIC slider U DICT U DICTS U DICTHR or U DICTHC 3 analyzer U AN 2 revolving nosepiece for DIC U D6RE U D7RE or U D7RES In order for reflected fluorescence to be effective in the simultaneous observation insert the U...

Страница 22: ...ot correctly engaged in light path Make sure that revolving nosepiece clicks properly into place Fluorescence mirror unit is not correctly engaged in light path Engage fluorescence mirror unit correctly in light path Field iris diaphragm is set too small Center aperture iris diaphragm and field iris diaphragm and then fully enlarge field iris diaphragm 11 ND slider is not stopped at click position...

Страница 23: ... 2 U excitation Narrow band U FUN Transmittance Wavelength nm 6 B excitation Narrow band U FBN Transmittance Wavelength nm 3 V excitation Narrow band U FVN Transmittance Wavelength nm 7 G excitation Wide band U FGW Transmittance Wavelength nm 4 BV excitation Wide band U FBVW Transmittance Wavelength nm 8 Y excitation Wide band U FYW Transmittance Wavelength nm ...

Страница 24: ...ion Wide band U FBWA Transmittance Wavelength nm 14 For cyan fluorescent protein CFP U FCFP Wavelength nm Transmittance 11 IB excitation color separation Narrow band U FBNA Transmittance Wavelength nm 15 For green fluorescent protein GFP U FGFP Wavelength nm Transmittance 12 G excitation color separation Wide band U FGWA Transmittance Wavelength nm 16 For yellow fluorescent protein YFP U FYFP Wave...

Страница 25: ...avelengths of 365 366 404 7 435 546 1 and 577 0 579 1 nm In addition light beams having wavelengths of 334 2 and 490 nm at rather low luminance are also applicable to fluorochrome emission Typical Example of Emission Spectrum of Ultrahigh Vacuum Mercury Burner Spectrum 17 For red fluorescent protein RFP U FRFP Transmittance Wavelength nm 18 For mCherry U FMCHE Transmittance Wavelength nm ...

Страница 26: ...ut control box For acquisition of mirror unit position information Lamp housing for mercury burner Lamp housing for 100 W mercury burner U LH100HG Apo lamp housing for 100 W mercury U LH100HGAPO Mercury burner USH 103OL OLYMPUS Power supply unit U RFL T Mercury burner DC 100 W ultrahigh pressure mercury burner USH 103OL OLYMPUS Life 300 hours 2 hour ON 30 min OFF cycle Operating environment Indoor...

Страница 27: ...for clamping the screws inside the illuminator to retain the performance have your Olympus representative conduct this work Be sure to insert the sliders in the orientations shown in the diagram Otherwise they cannot be fitted in click positions and engaged correctly in the light path To prevent fire install the lamp housing with the heat radiating fins positioned on the top and with sufficient sp...

Страница 28: ...4 Otherwise the fluorescence mirror unit may be mounted being tilted Two more fluorescence mirror units can be mounted in the turret positions on the left and right in the current condition Write down the turret position numbers and fluorescent mirror unit names for use when you insert the indication sheets later 5 Mount other necessary fluorescence mirror units in the same way as above Do not rem...

Страница 29: ...h burner to the original position and tighten the clamping screw Align the external edges of the lamp housing with those on the socket section and push the lamp housing straight down ward Attach the lamp housing on the lamp housing installation po sition of the microscope system so that the heat radiating fins face upward To prevent a fire hazard reserve ample spaces above below and on the rear of...

Страница 30: ...ely Place each filter by aligning the directions of the arrows located on the side surfaces toward the absorption filter frame and the main body respectively Fit the holder ring driver into the groove of the holder ring driver and fix it by rotating the filter holder ring If the filter holder rings may exceed the absorption filter frame or the main body depending on the filter thickness re attach ...

Страница 31: ... inspections are charged after expiration of the warranty period If you have any questions please contact Olympus Check results Date Check items 1 More than 8 years have passed since original purchase or the total power ON time has exceeded 20 000 hours 2 Illumination flickers when you move the lamp cable or lamp housing 3 Lamp cable is unusually hot to the touch 4 Scorching or burning odor is pro...

Страница 32: ...ng Table 1 Certified Cord A power supply cord should be certified by one of the agencies listed in Table 1 or comprised of cordage marked with an agency marking per Table 1 or marked per Table 2 The fittings are to be marked with at least one of agencies listed in Table 1 In case you are unable to buy locally in your country the power supply cord which is approved by one of the agencies mentioned ...

Страница 33: ...Cables BASEC BASEC HAR 10 10 30 N V KEMA KEMA KEUR HAR 10 30 30 SEMKO AB Svenska Elektriska Materielkontrollanstalter SEMKO HAR 10 10 50 Österreichischer Verband für Elektrotechnik ÖVE ÖVE HAR 30 10 50 Danmarks Elektriske Materialkontroll DEMKO DEMKO HAR 30 10 30 National Standards Authority of Ireland NSAI NSAI HAR 30 30 50 Norges Elektriske Materiellkontroll NEMKO NEMKO HAR 10 10 70 Asociacion E...

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