Olympus BX-RFA, Instruction Manual

Page 1: ...tion manual is for the Olympus Reflected Fluorescence System To ensure the safety obtain optimum performance and to familiarize yourself fully with the use of this system we recom mend that you study...

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Page 3: ...EOUS FLUORESCENCE OBSERVATIONS 5 TROUBLESHOOTING GUIDE 6 SPECTRAL CHARACTERISTICS OF FILTERS 7 SPECIFICATIONS 4 5 6 7 8 15 16 17 18 22 23 1 General Precautions for Observation 8 2 Selecting the Fluore...

Page 4: ...ted Light Nomarski Differential Interference Contrast DIC Observation 40 42 4 3 Reflected Light Simple Polarized Light Observation 42 5 OPTICAL CHARACTERISTICS UIS2 UIS Series for Reflected Light Obse...

Page 5: ...struments Handle it with care and avoid subjecting it to sudden or severe impact 2 The ultrahigh pressure mercury burner used should be the USH 103OL DC burner mfd by USHIO Inc or the HBO103W 2 burner...

Page 6: ...s U 25ND6 U 25ND25 U 25ND50 Warning against high temperature Warning against high voltage 1 Getting Ready 1 This manual pertains only to the reflected fluorescence system Before using this system toge...

Page 7: ...value set the main switch to OFF and wait for more than 10 minutes before replacing the mercury burner Page 33 Unlike electric bulbs the mercury burner seals high pressure gas inside If it continues t...

Page 8: ...knob Page 12 Shutter OUT Shutter IN Aperture iris diaphragm knob Page 13 Collector lens focusing knob Page 14 Burner centering knobs Page 14 Mirror focusing screw Page 15 On the rear of the lamp hous...

Page 9: ...ot recommended to open a fluorescence mirror unit It is recommended that you use the U MF2 dummy filter unit which does not contain a filter when making your original fluorescence unit Page 32 Blank i...

Page 10: ...ages 8 to 11 Center the mercury burner Page 14 or 15 Main switch Place the specimen on the stage Specimen holder X Y axis knobs Engage the fluorescence mirror unit matching the specimen in the light p...

Page 11: ...7 Make a photocopy of the observation procedure pages and post it near your microscope...

Page 12: ...s 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 fading...

Page 13: ...bserving only the B excitation stain when using B excitation stain with TRITC or Texas Red Mirror Unit Name Meaning For color separation U M N I B A 2 Excitation U V BV B IB G IG or IY Bandwidth SW Su...

Page 14: ...ht DIC UPlanSApo 4X 10X 2 20X 20X O 40X 2 60X W 60X O 100X O CFP U MCFPHQ DM450HQ BP425 445HQ BA460 510HQ For ECFP GFP U MGFPHQ DM485HQ BP460 480HQ BA495 540HQ For EGFP YFP U MYFPHQ DM505HQ BP490 500H...

Page 15: ...ition The mercury burner cannot be reignited until the mercury vapor has cooled down and liquefied Before re igniting a mercury burner wait for about 10 minutes after the last time it was turned off F...

Page 16: ...iris image comes at the center of the field of view 7 While pushing in the field iris diaphragm knob enlarge the field iris diaphragm until the field iris image inscribes the field of view If eccentr...

Page 17: ...the aperture iris diaphragm knob to minimize the aperture iris diameter 8 Pull out the field iris diaphragm knob to minimize the field iris dia phragm Now the aperture iris image should be visible on...

Page 18: ...osepiece to engage the empty position the objective cap should be removed in the light path 5 Pull out the field iris diaphragm knob to minimize it and push in the aperture iris diaphragm knob to enla...

Page 19: ...arc image using the Allen screwdriver in these holes Fig 5 8 Mounting the ND Filters Specimen color fading can be delayed by reducing the excitation light intensity with ND filters Use the ND filters...

Page 20: ...d fluorescence to be effective in the simultaneous observation insert the analyzer U AN or U AN360 3 into the analyzer inlet slot above the dichroic mirror on the illuminator Do not insert the U ANT a...

Page 21: ...or not evenly illuminated Objective is not correctly engaged in light path Make sure that revolving nosepiece clicks properly into place Fluorescence mirror unit is not correctly engaged in light path...

Page 22: ...n Narrow band 4 BV excitation Wide band 5 BV excitation Narrow band 6 B excitation Wide band U MWU2 U MNU2 U MNV2 U MWBV2 U MNBV2 U MWB2 Transmittance Wavelength nm Transmittance Wavelength nm Transmi...

Page 23: ...rwide band 9 IB excitation Wide band 10 IB excitation Narrow band 11 G excitation Wide band 12 G excitation Narrow band U MNB2 U MSWB2 U MWIB3 U MNIB3 U MWG2 U MNG2 Transmittance Wavelength nm Transmi...

Page 24: ...tation color separation Wide band 18 IB excitation color separation Narrow band U MSWG2 U MWIG3 U MWIY2 U MNUA2 U MWIBA3 U MNIBA3 100 400 600 800 0 BA510 550 BP460 495 DM505 100 400 600 800 0 BP470 49...

Page 25: ...BP535 555HQ BA570 625HQ DM565HQ 19 G excitation for color separation Wide band 20 G excitation for color separation Narrow band 21 For cyan fluorescent protein CFP 22 For green fluorescent protein GFP...

Page 26: ...t 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...

Page 27: ...uorescence Phase contrast Reflected light brightfield Reflected light darkfield Reflected light DIC Reflected light simplified polarization Transmitted light Reflected fluorescence Reflected fluoresce...

Page 28: ...each filter so that it does not fit inside the holder ring If you perform transmitted light observation or you do not want to use a filter mount the provided light shield plates having the same size...

Page 29: ...comes at the deep Otherwise the filters will not be set in the correct positioning When the 6 position filter slider near the rear panel is used avoid using the interference type or color glass type...

Page 30: ...icker position Make sure that the sticker does not deviate from the specified circular area Otherwise the slide will be caught in motion 4 Gently insert the filter slider from the right of the analyze...

Page 31: ...n 1 Insert the provided adjustment knobs into the two adjustment screw holes near the front panel and move the two sides of the rectangle to the desired position by turning the knobs 2 Insert the adju...

Page 32: ...llen screwdriver Operation Observing a Double Stained Specimen Due to its own characteristics the G exci tation has a narrower intensity control range than the U and B excitation The intensity control...

Page 33: ...f shorter wavelengths near blue Front Observer Observation of Triple Stained Specimen The operation is basically similar to the double stained specimens but fluorescence mirror units for triple staini...

Page 34: ...have your Olympus representative conduct this work ASSEMBLY 9 1 Assembly Diagram NOTES Parts marked with can be attached only to the BX URA2 universal illuminator Be sure to insert the sliders in the...

Page 35: ...unit to be mounted so that the model name inscription on the side is upside down align it with the mount dovetail and insert all the way into the insertion position Tighten the clamping screw firmly I...

Page 36: ...unit frame Dimensions of Optical Parts Barrier filter Excitation filter Dichroic mirror 26 0 1 0 3 x 38 0 1 0 3 mm thickness 1 0 05 mm When replacing the dichroic mirror take special care not to stai...

Page 37: ...r holding screws 4 Attach the positive pole of a specified mercury burner to the fixed mount on the upper side and the negative pole to the mount on the lower side Be sure to use the USH 103OL mfd by...

Page 38: ...kering is likely to increase It is therefore recommended to replace the burner according to the purpose of observation This value assumes light cycles composed of 2 hours of lighting and 30 minutes of...

Page 39: ...input power supply match the requirements inscribed on the rating plate 100 V systems can be used with voltages in the 100 to 120 V range and 200 V systems can be used with voltage in the 220 to 240 V...

Page 40: ...or unit Objective Light shield tube DIC prism U AN U AN360 3 U 25ND6 2 U 25ND25 2 U 25ND50 2 U 25LBD U 25IF550 U 25Y48 U 25FR U 25L42 U RCV Mercury lamp housing U LH100HG U LH100HGAPO Halogen lamp hou...

Page 41: ...ield Tube Fig 19 The light shield tube must be used with darkfield observation using DF mirror unit 1 Remove the turret 2 Place the light shied tube in the reflected illuminator so that the position i...

Page 42: ...e Iris Diaphragm Fig 21 1 Engage the mirror unit BF in the light path by turning the turret then open the shutter 2 Rotate the revolving nosepiece to engage the 10X objective then place a highly flat...

Page 43: ...tfield BF U MBF3 Adjust as required Reflected light darkfield DF U MDF3 Must be open Applications U 25FR Frost filter To eliminate uneven illumination U 25LBD Color temperature conversion filter To co...

Page 44: ...in use Fig 24 1 Selecting the Light Path for Observation Fig 24 1 Rotate the turret to engage the BF mirror unit in the light path When the U MDIC3 DIC mirror unit is mounted in the turret engage the...

Page 45: ...observation with good con trast is possible in the most sensitive gray colors If the background color is sensitive magenta even a minor optical retar dation can be observed as a color change 1 Rotate...

Page 46: ...Light Simple Polarized Light Observation To prepare for simple polarized light observation using the reflected illuminator perform the operations in paragraph Selecting the Light Path in section 4 2 R...

Page 47: ...39 12 5X 870 17 6 2 5X 0 08 10 7 4 19 25X 220 8 8 25X 220 10 6 5X 0 15 20 0 2 24 50X 59 4 4 50X 59 5 3 10X 0 30 11 0 1 12 100X 15 2 2 100X 15 2 65 20X 0 45 3 1 0 0 75 200X 5 2 1 1 200X 5 2 1 33 50X 0...

Page 48: ...e top of specimen and the front lens of objective Number of aperture NA Important figure determining the objective characteristics resolution focal depth and bright ness Resolution Increases in propor...

Page 49: ...correctly Center mercury burner Frost filter is not engaged in light path Engage frost filter in light path Filter is not in click position Push filter until it clicks properly c Image glares Apertur...

Page 50: ...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 fit...

Page 51: ...Authority of Ireland NSAI Norges Elektriske Materiellkontroll NEMKO Asociacion Electrotecnica Y Electronica Espanola AEE Hellenic Organization for Standardization ELOT Instituto Portages da Qualidade...

Page 52: ...ny 3500 Corporate Parkway P O Box 610 Center Valley PA 18034 0610 U S A One Corporate Drive Orangeburg NY 10962 U S A 491B River Valley Road 12 01 04 Valley Point Office Tower Singapore 248373 31 Gilb...