facs diva training garvan mlc flow cytometry facility

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Garvan MLC flow Cytometry Facility guide to using Facs DiVa

TRANSCRIPT

MLC Flow Cytometry Facility

BD FACS DiVa

Training

Flow Cytometry

Login

• Windows Login • User - Administrator

• Password – BDIS or bdis

• DiVa Login• Username - 3 letters first name + 3 letters last name

• Password – as user name

YOUR DISCIPLINE HERE

DiVa Overview

DiVa Overview

Browser

Window

DiVa Overview

Browser

Window

Cytometer

Window

DiVa Overview

Browser

Window

Cytometer

Window

Worksheet

Window

DiVa Overview

Browser

Window

Cytometer

Window

Worksheet

Window

Acquisition

Dashboard

DiVa Overview

Browser

Window

Cytometer

Window

Worksheet

Window

Acquisition

Dashboard

Inspector

DiVa Overview

Browser

Window

Cytometer

Window

Worksheet

Window

Acquisition

Dashboard

Inspector

DiVa Overview

Browser

Window

Cytometer

Window

Worksheet

Window

Acquisition

Dashboard

Inspector

File menu

DiVa Windows -Browser

Organises

data in a

hierarchical

and ordered

manner

DiVa Windows - Browser

DiVa Windows - Browser

DiVa Windows - Cytometer

Status - instrument errors

( turns red)

Parameters – controls PMT

amplification and signal

collection

Threshold – controls triggering

Laser – Laser delays and

scaling

Compensation – acquisition

defined compensation

matrix

Ratio - ???

DiVa Windows - Cytometer

DiVa Windows -Cytometer

Parameter

name

DiVa Windows - Cytometer

DiVa Windows - Cytometer

DiVa Windows - Cytometer

Worksheets• Global worksheet

• Is not linked to specific data source

• Reloads data on tube selection

• Used when batching

• Different global worksheets can be linked to different

specimens

• Sheet • Is linked to data

• Does not refresh upon tube selection

• Compensation • Is linked to compensation tubes

• Must be used to perform acquisition compensation

Worksheets

Visualising Signal Data

Dot

plot

Histogram

Density

plot

Contour

plot

Statistical

measures are

also used to

identify

changes

DiVa Windows - Acquisition

Dashboard

DiVa Windows - Acquisition

Dashboard

DiVa Windows - Acquisition

Dashboard

DiVa Windows - Acquisition

Dashboard

DiVa Windows – Inspector

DiVa Windows – Inspector

DiVa Windows – Inspector

DiVa Windows – Inspector

Creating a New Experiment

Creating a New Experiment

Creating a New Experiment

Compensation Workflow

Compensation Workflow

Compensation Workflow

Using the Layout editor

Using the Layout editor

Using the Layout editor

Fluidics Startup/Shutdown

Canto I, Canto II and Aria only

Fluidics Startup/Shutdown

Canto I, Canto II and Aria only

Fluidics Startup/Shutdown

Canto I, Canto II and Aria only

Acquiring data

• Activate tube to collect data

• Hit acquire

• Check stopping gate and numbers

• Hit record

• Hit remove tube

• Remove tube and flick aspirator arm to left

• repeate

Exporting Data

Exporting Data

Exporting Data

Cleaning Instrument

1. Run 10 % fresh bleach soln. – 5 mins*

2. Run 4 % extran – 5 mins

3. Run distilled water – 5 mins

4. Shutdown if no other users in the day

* For Biohazardous samples

Troubleshooting

Troubleshooting

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