Olympus 1X71 User Manual Download | Manualshive

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Selecting the right fluorochrome/filter set

Position #1
49000 - ET - DAPI

Exciter 350/50x

beamsplitter 40011

3

Emitter ET460/50m

AT350/50x

T4001p

ET460/50m

Fluorochrome

EX

EM Use

Alexa Fluor 350TM

346 442 Recommended

Coumarin

384 470 Recommended

DAPI

359 461 Recommended

DyLight 350

352 435 Recommended

Hoechst 33258

352 461 Recommended

LysoTracker Blue/Me0H

373 425 Recommended

550

Wavelength (nm)

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Summary of Contents for 1X71

Page 1: ...ember 2014 UCL Institute of Child Health User guide Olympus 1X71 Dr Bertrand Vernay Light Microscopy Facility Manager Wellcome building Office W2 09 Tel Office 42224 Tel Mobile 07582249337 Email b vernay ucl ac uk ...

Page 2: ...User Guides Transmitted Light Epifluorescence Image Capture page 6 Halogen Lamp Operation page 7 Kohler Illumination page 8 Adjusting the Objective Correction Collar page 9 Prior Lumen200 Metal Halide Lamp Operation page 10 Image Capture with HClmage page 11 Selecting the Right Fluorochrome Filter Set page 11 DAPI page 12 Endow GFP EGFP Bandpass page 13 DsRed TRITC Cy3 page 14 Cy5 page 15 Cy7 page...

Page 3: ...rge for all other users towards the cost of the consumables is expected Prof Sowden team has priority over other users Users must always record their activity in the Log book Problem s with the microscope should be reported as soon as they are noticed Olympus customer support service http www olympus co uk microscopy The system is not covered by a maintenance contract Olympus requires a PO number ...

Page 4: ...0LPXR ET700 75m 5 Cy7 ET710 75x T760LPXR ET810 90m 6 empty brightfield Camera see p16 Hamamatsu ORCA R2 CCD Camera with HClmage Capture Software Fluorescence illumination see p16 Prior Lumen 200 Metal Halide Light Source 2000 hours bulb Pixels to Microns calibration 5x objective binning 1x1 1 pixel 1 88 um 10x objective binning 1x1 1 pixel 1 03 um 20x objective binning 1x1 1 pixel 0 514 um 40x obj...

Page 5: ...e to do so is likely to result in damage to the bulb 1 Prior Lumen 200 module on 2 Prior Lumen 200 intensity knob 0 3 Light Path selector lever on Ocular 4 Correct filter cube in position 5 Fluorescence shutter open Image Capture 1 Start Camera controller press until LED turns green 2 Computer on Login Jane Password Ja e 3 HClmage software open 4 Light Path selector lever on Camera 5 Correct trans...

Page 6: ...e light intensity control knob 1 is in the MIN minimum intensity position on the TH4 module 3 Set the main switch 2 to I ON on the TH4 module 4 On the microscope front press the transmitted light ON OFF button 6 so that the button is illuminated 5 Adjust the brightness with the light intensity control knob 5 6 To turn OFF set the transmitted light ON OFF button 6 to OFF ...

Page 7: ...Set the light intensity control knob 5 to the MIN minimum intensity position on the microscope frame 2 Set the light intensity control knob 1 to the MIN minimum intensity position on the TH4 module 3 Set the main switch 2 to 0 OFF on the TH4 module ...

Page 8: ...ing the specimen into focu s 5 Using the field iris diaphragm lever 3 completly close the field iris diaphragm 6 Rotate the condensor height adjustment knob 4 to bring the field iris diaphragm image into focus 7 Center the field iris diaphragm 3 using the condenser centering knobs 5 8 Open the field iris diaphragm 3 until its image reach the limits of the field of view adjust the centering if nece...

Page 9: ...can be obtained by judging the image resolution and contrast When a satisfactory image is not obtain after focusing 1 Rotate the correction collar to the left and right refocus each time and compare the images 2 Then rotate the collar in the direction yielding a better image rotate the correction collar to the left and right refocus each time and compare the images 3 Repeat this cycle until the po...

Page 10: ...ational temperature 5 Warning Do not power down the unit within 30 mins of power up This may re duce the effective lifetime of the bulb Shutting down the Lumen The following warnings apply as damage to the bulb may result if instructions not followed 1 Warning Do not shut the unit down within 30 minutes of powering up the unit 2 Warning After shutting down the unit allow 30 minutes before re power...

Page 11: ...cking on Auto Expose view the intensity distribution in the histogram 5 Check Sat saturation in the histogram of the Image Display to guard against image saturation Saturated pixel are indicated in Red Yellow indicates pixels ap proaching saturation 6 Adjust camera exposure and gain settings as necessary 7 Click Abort 8 Click Capture1 to acquire an image 9 Click the Save icon to save the image in ...

Page 12: ...tter 400113 Emitter ET460 50m AT350 50x T4001p ET460 50m Fluorochrome EX EM Use Alexa Fluor 350TM 346 442 Recommended Coumarin 384 470 Recommended DAPI 359 461 Recommended DyLight 350 352 435 Recommended Hoechst 33258 352 461 Recommended LysoTracker Blue Me0H 373 425 Recommended 550 Wavelength nm ...

Page 13: ...05 512 Alternative Calcein 494 517 Alternative Calcium GreenTM 1 506 531 Alternative Cy2TM 489 506 Alternative DiO 484 502 Alternative DyLight 488 492 517 Alternative EGFP 488 507 Alternative Emerald GFP 489 510 Alternative FAM 492 518 Alternative FITC 490 525 Alternative Fluo 4 494 516 Alternative GFP 488 507 Alternative MitoTracker Green FM MeOH 490 516 Alternative mWasabi 493 509 Alternative Or...

Page 14: ...0 603 Recommended Ethidium homidimer 1 DNA 527 617 Recommended Propidium Iodide 536 617 Recommended Resorufin 571 585 Recommended Alexa Fluor 568TM 78 603 Alternative Cy3TM 552 570 Alternative Rhod 2 540 576 Alternative TAM RA 555 580 Alternative Tetramethylrhodamine isothio cyanate 555 580 Alternative TRITC 555 580 Alternative I 50 100 10 40 30 20 60 80 70 90 0 ____________________ 450 500 550 60...

Page 15: ...66 Recommended Allophycocyanin APC 630 660 Recommended Atto 647N 644 669 Recommended Cy5TM 649 670 Recommended DiD 644 665 Recommended Draq5 647 683 Recommended DyLight 649 652 667 Recommended MitoTracker Deep Red 633 MeOH 644 665 Recommended Nile Blue 631 660 Recommended SYTO 60 652 678 Recommended TO PRO TM 3 642 661 Recommended 750 800 ...

Page 16: ...ter ET810 90m 100 90 80 70 60 I 50 s t 40 30 20 10 0 ______ 600650 700 750 800 850 900 Wavelength nm ET710 75x T7601pxr ET810 90m Fluorochrome EX EM Use Alexa Fluor 750TM 752 779 Recommended Cy7TM 743 767 Recommended DiR 748 780 Recommended DyLight 750 751 772 Recommended ...

Page 17: ...HClmage Live ...

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