Olympus Widefield Fluorescence System


Location: T-lab, level10, Microscope Room, Room 9


Microscope Specifications for Olympus Widefield Fluorescence System
 
 
 

Light Source               Metal Halide Lamp

Microscope Body       Olympus IX81 Inverted Microscope

Beam splitters and filters

 

GFP

TRITC

DAPI/Hoechst/AMCA

CY5

 

Excitation Filters

470/40

530/20

330/55

 

 

Dichroic Mirrors

505

570

400

 

 

Emission Filters

510/50

575

420

 

 

 

Objectives

Model

Immersion

N.A.

W.D. (mm)

Remarks

U Plan FLN 10XPH (Air)

Nil

0.30

10

 

LUC Plan FLN 20XPH (Air)

Nil

0.45

6.6 – 7.8

Long Working Distance

LUC Plan FLN 40XPH (Air)

Nil

0.60

3.0 – 4.2

Long Working Distance

LUC Plan FLN 60X (Air)

Nil

0.70

1.5 – 2.2

 

U Plan SAPO 60XW (Water)

Water

1.20

0.28

 

U Plan SAPO 100XO (Oil)

Oil

1.40

0.13

 

 

Camera                       CoolSnap liquid cooled CCD (EZ1394) Digital Monochrome Camera

Operation system and software

WinXP OS

2GB RAM, 500GB System Disk, DVD-RW, VGA card

Operation Software: MetaMorph

Misc. Accessories

Motorized stage with Linear Encoder (Ludl)

6-well petri dish adapter with heated cover

Incubator: Temperature, Humidity, CO2 control

ZDC: Z focus drift compensator

Phase contrast for 10X/20X

DIC for 10x/20x/40x

SOP for Olympus Widefield Fluorescence System

1. Getting Started

 

Step 1: To switch on the system and PC.

a)  Switch on switch labelled "Main Switch"

  

 

b) Switch on Mercury Lamp on top of the rack.

- Wait until the Light Indicating Burner On is stable.

- Caution!! If it's not stable for 1 minute, the lamp needs to be replaced. Please contact microscope staff.

 

c) Switch on Transmitted light ON-OFF button

 

 

d) Switch on the CO2 supply with the label of ‘LIVE CELL’ at the side of the wall

Caution!! CO2 supply shared with Nikon Biostation, be sure that you are switching on the right switch

 

d) Turn on PC.

 

e) Click on the Metamorph Software icon and proceed with imaging procedures.

     

                                                          

 

Step 2:

a) Select the required Neutral Density filters.

 

 

b) Adjust the brightness.

                - Light intensity buttons

 

c) Engage a 10X objective in the light path.

- Objective switching button

 

d) Select the bright field observation and select the (observation) light path.

 

 

Step 3:

a) Place the specimen on the stage.

 

 

b) Bring the specimen in focus.

- Focus adjustment knob

- Adjust the brightness.

 

 

e) Adjust the light path.

-  Adjust the aperture iris and field iris diaphragms.

 

 

Step 4:

a) Engage the objective to be used in the light path and bring the specimen in focus.

- Objective switching button

- Focusing knob (dial)

- Focus adjustment knob

 

Caution: Ensure the required objective is placed in the correct objective pocket in the microscope.

 

Caution: Clean the oil objectives or wipe the water lens after use. Do not switch back and forth between Oil objectives and air objectives without wiping the oil away from the oil objectives first.

Step 5:

 

Start observation.

 


 
 

2. Multi-dimensional acquisition 

 

Goal: To acquire multi-modal images with combination of different wavelengths, optical sections, multiple stage positions etc.

 

Overview of procedures:

 

1. Choose the modes e.g. time, Z, wavelengths etc to include in experiment

2. Choose a subdirectory with enough space for the experiment

3. Set up dataset name. This will be a *.ND file managing the image files comprising of TIFF and stacks

4. Set up individual tabs

5. Confirm exposure times and window positions using Snap or Preview alternating among the different wavelengths

6. Click "Acquire" to start experiment

7. Press ESC if experiment is to be terminated before completion

 

 

 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 

3. Switching off the system

 

a) Exit from Metomorph software

 


b) Switch off Transmitted light ON-OFF button

 


c) Switch off Mercury Lamp on top of the rack.

 


c) Switch off the Control Box

 


d) Switch off the CO2 supply with the label of ‘LIVE CELL’ at the side of the wall

Caution!! CO2 supply shared with Nikon Biostation, be sure that you are switching on the right switch

 


d) Turn off PC and collect your sample

Caution!! Clean the chamber if there is a spillage, be extra careful when cleaning the lens. 

 
 For further enquiries, please contact mbiwpy@nus.edu.sg
 

 

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