background image

 

p.

 

12

 

 

Single

 

image

 

acquisition

 

 

In

 

the

 

Acquisition

 

Mode

 

tab

 

select

 

the

 

preferred

 

zoom

 

(alternatively

 

you

 

can

 

easily

 

position

 

the

 

cropping

 

frame

 

to

 

zoom

 

in

 

via

 

the

 

crop

 

tool

 

 

below

 

the

 

image

 

in

 

the

 

Dimensions

 

tab)

 

 

 

Scan

 

one

 

frame

 

to

 

check

 

position.

 

 

Check

 

all

 

tracks

 

in

 

the

 

Channel

 

tab,

 

In

 

the

 

Acquisition

 

Mode

 

tab

 

click

 

 

to

 

set

 

the

 

frame

 

size.

  

 

Set

 

scanning

 

speed

 

to

 

6

 

 

Set

 

averaging

 

number

 

to

 

2

4

 

(according

 

to

 

the

 

SNR)

 

 

Press

 

Snap

 

  

in

 

the

 

start

 

buttons

  

 

Save

 

the

 

image:

 

press

 

 

in

 

the

 

Images

 

and

 

Documents

 

tab

 

(right

 

hand

 

side

 

of

 

the

 

window)

 

and

 

save

 

in

 

the

 

directory:

  

D:\Users

 

data\PI

 

name\MM

YYYY\User

 

name\YYYY

MM

DD

 

 

Multidimensional

 

acquisition

  

 

Z

 

stack

 

 

In

 

the

 

Multidimensional

 

Acquisition

 

options

 

below

 

the

 

start

 

buttons

 

check

 

Z

 

stack

;

 

a

 

new

 

tab

 

(

Z

stack

)

 

will

 

appear

 

under

 

the

 

Multidimensional

 

Acquisition

 

Section

.

 

 

Set

 

the

 

interval:

 

press

 

Smallest

 

for

 

the

 

system

 

recommendation

 

(Nyquist

 

criterion)

 

 

With

 

one

 

channel

 

checked

 

and

 

In

 

the

 

Acquisition

 

mode

 

tab:

 

Frame

 

size

   

512X512

 

Scanning

 

speed

 

8

  

Averaging

 

number

 

1

 

Summary of Contents for LSM 700

Page 1: ...2 Objectives 2 Filter turret 2 Laser lines 3 Light Path 3 Start up 4 Sample mounting and viewing 5 Software 5 Sample viewing 6 Acquisition 8 LSM Channel mode confocal setting 8 Configuration 8 Acquis...

Page 2: ...pectral data point Hardware The microscope The LSM 700 is attached to an upright microscope Axio Imager Z2 Objectives Position Objective lens Magnification NA coverslip Working distance Immersion Parf...

Page 3: ...mission passes through the pinhole aperture and the out of focus light is eliminated 5 6 The variable secondary dichroic beamsplitter VSD is coated with a variable coating that between 420nm 630nm acc...

Page 4: ...ng order In the power strips turn on switches 1 2 Turn on the stage controller 3 Rotate clockwise the silver key in the lasers electronic unit 4 Turn on the computer and log in User name multilabs Pas...

Page 5: ...Light switch between the ocular and the confocal Via the touch pad 1 Choose the preferred objective 2 Choose the proper filter set and open the TL RL shutter Software Log into BookitLab and activate...

Page 6: ...tage to I II or III according to the objective lens see objective lens info on touchpad If you are acquiring transmitted channel set up Kohler illumination 1 Bring the sample into focus 2 Close the fi...

Page 7: ...p 7 2 5 3 4...

Page 8: ...ocal Configuration It is highly recommended to seek BCF staff s advice for new fluorophores exact name or new combinations Choose one Load a saved configuration from the Experiment Manager Reuse setti...

Page 9: ...ThermoFisher Chroma 1 Laser lines 2 Main dichroic mirror 3 PMTs PMT1 and PMT2 4 Variable secondary dichroic beamsplitter VSD 5 Emission Filter selection 6 Emission spectrum with Graphic representatio...

Page 10: ...logy experiments Frame size 512X512 Speed 8 Averaging Method mean in most cases in low signal with low noise Sum method may be useful Number 1 Mode frame for fixative sample for live cells and in case...

Page 11: ...lack level black blue pixels in the image o Set the PMT gain to 550 600 o Set the Laser it is recommended to use the lowest as possible and to increase gain master o In case of low signal with high la...

Page 12: ...ng to the SNR Press Snap in the start buttons Save the image press in the Images and Documents tab right hand side of the window and save in the directory D Users data PI name MM YYYY User name YYYY M...

Page 13: ...uch as brightness correction over z Time series In the Multidimensional Acquisition options check Time series a new tab Time Series will appear under the Multidimensional Acquisition Section Define nu...

Page 14: ...mended option the process can be configured in the processing main tool tab In the Acquisition mode tab Frame size Press optimal Zoom It is recommended to set the zoom above 1 2 Scanning speed 7 Avera...

Page 15: ...After a single scan in the Regions tab choose one of the ROI tools and draw on the acquired image the dimensions can also be set manually in the Width Height textboxes Define which process will be per...

Page 16: ...before moving to high magnification with small field of view Add a position by pressing Positions below the image container and clicking on the image container that was just acquired at the current st...

Page 17: ...lose the ZEN software Log off from your account in the BookItlab window Check if another user has a reservation if yes verify his her arrival If you are the last user for the day or the next reservati...

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