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Last updated on Mar 2022 

3.1 Acknowledgements 

If you use the TLL Microscopy and Imaging facility and/or have been trained or assisted any 
of the bioimaging facility members in your research, then this should be acknowledged 
appropriately in your publications and presentations. 

4. Operation Procedures 

Every confocal system in TLL Bioimaging Facility has its specific instructional manual which 
are found in every confocal rooms. Strictly adhere to the correct order of operation for all 
system. Failure to do so will result in disciplinary action from the facility. Any issues 
encountered during the operation of the system are advised to seek help from the 
Bioimaging facility.  

Modification, exchange or removal of components beyond this operational manual is strictly 
prohibited and is only carried out by the manufacturer, Bioimaging facility or by experienced 
users approved by bioimaging facility. During operation of laser microscopy system, do not 
look into the laser beam directly as they are all Class 3b and Class 4 lasers.  

 

4.1 Switching ON Protocol  

(a)

 

SIGN IN the Logbook and record your START-TIMING. 

(b)

 

Switch on the Master switch on the wall 

(1)

 & PC

 (2)

. Wait until windows icon pops 

up. 

(c)

 

Switch on Stage controller box 

(3)

 & CBH controller 

(4)

.  

(d)

 

(Optional)

 Turn on the Metal Halide lamp 

(5)

, if needed. 

(e)

 

Turn on the Touch screen panel power button at the back 

(6)

 and wait for 5s to light 

up. 

(f)

 

Switching ON Lasers, 

 

To use 405/488/561/640nm, switch on the power button on the main 
combiner 

(7)

, then turn the key to “ON” 

(7A)

 and finally turn on the 

individual lasers that you need. 

 

To use 445/514nm, switch on the power button on the sub combiner 

(8),

 

then turn the key to “ON” 

(8A)

 and finally turn on the individual lasers that 

you need. 

(g)

 

(Optional)

 Turn on the incubator 

(9)

 if required. It takes 10-15mins to stabilize. 

(h)

 

Mount your slide onto the sample holder. If you need Live cell Imaging, turn on the 
OkaLab incubator and the facility CO

2

 (10)

. It takes 10mins to stabilize. If you require 

help mounting the stage incubator holder onto the stage, please request help from 
the TLL bioimaging department.  

(i)

 

Log in with your TLL user account and d

ouble click on “FV31S

-

SW” to load the 

software. 

1st offence 

A warning will be issued along with the reminder of the rules. 

2nd offence 

A second warning will be issued and your respective PI will be notified. 

3rd offence 

Banned from using any of the facility’s microscope for 2 weeks.

 

Summary of Contents for FV3000

Page 1: ...oking and System Access 4 3 1 Acknowledgements 5 4 Operation Procedures 5 4 1 Switching ON Protocol 5 4 2 Objective lens 6 4 2 Software Initiation 7 4 3 Setting up directory to save images 8 4 4 Laser...

Page 2: ...Immersion Oil Modern immersion oil has no known hazards to human beings so far yet they can cause discomfort to a person if the immersion oil has been left on skin for too long or inhaled Therefore pl...

Page 3: ...taff before they are allowed to access the confocal system 2 1 To arrange a training session Please email Bioimaging Facility bioimaging tll org sg for a training session Fill in the necessary particu...

Page 4: ...er 6pm Sat Sun Public holidays If users require more slots in a particular week they can book on weekdays during non peak hours after 6pm on weekends Sat and Sun and on public holidays If extra time i...

Page 5: ...RT TIMING b Switch on the Master switch on the wall 1 PC 2 Wait until windows icon pops up c Switch on Stage controller box 3 CBH controller 4 d Optional Turn on the Metal Halide lamp 5 if needed e Tu...

Page 6: ...t to 10x objective or lower then click YES Cleaning the stage will take some time b Otherwise click NO to skip cleaning 4 2 Objective lens Objective lens Immersion media 1 25x 0 04 Plan Apochromat Air...

Page 7: ...n the stage mechanism This adds time to the initialization process Do not place anything on the stage mechanism until the initialization process is complete If you are not the first user of the week C...

Page 8: ...y to save all your images Please save in your folder You have several different options on where you can save your images Option 1 D or E Users shaalini create your own folder BPAE cells oir Do keep i...

Page 9: ...oduce intense background noise SD keep HV within 600 800 More than 800 will contribute to intense background noise Under PMT settings tab Mode Select VBF Average average a given number of scans to get...

Page 10: ...to zoom images only in the center Ideal zoom which satisfy the Nyquist limit can be achieved by zooming till the red arrow any zoom beyond it will result in poor resolution and result in more photoble...

Page 11: ...the laser power until red pixels disappear Increase Offset until a dusting of blue pixels in the background are observed Click LSM start to capture an image 4 7 How to do a Z stack for a 3D Image xyz...

Page 12: ...your timelapse experiment Interval FreeRun continues to capture images until it has completed your selected cycle You could always choose an interval of your choice 4 9 Reuse previous settings and ex...

Page 13: ...h the sample and return the stage to central position d Carefully clean the objective lens with Whatman lens tissue using 100 ethanol e Switch lens back to 10x 0 3 objective lens f Exit the FV31S SW s...

Page 14: ...ission please contact Bioimaging Group No stage movement Check the coarse fine movement setting near the focus and xy position knobs It is a physical button that has no electronic indication elsewhere...

Page 15: ...id white or coloured light sometimes red light might be indicative of error III Restart the main controller right at the bottom if none of the steps taken work especially if the machine has been left...

Page 16: ...and cannot be recovered Restart computer terminal Startup popup error code LAST_OPERATION_ NONCOMPLETED The last user did not shut down the program properly or the program encountered a fatal error du...

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