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Modern fluorescence imaging systems are complex instruments made of many optical, mechanical, and electronical components. The possibilities of misalignment, malfunction or failure of imaging instruments can come from different components. Therefore, regularly controlling the quality of such instruments is important.

Aim:

  • Ensuring the acquisition of reliable data and preventing corrupted data

  • Removing the bias introduced by the instrument

  • Knowing how the system performances evolve over time and perform quantitative microscopy

Material: 

Argo SIM QC slide (Manufactured by Argolight France. Purchased in Dec 2021). This slide is composed of different fluorescent patterns in 2D and 3D. Below is an overview of available fluorescent patterns and corresponding quality tests.

...

Quality Test

Corresponding Pattern

Patten Image

QC Frequency

Lateral resolution

Gradually spaced lines

Image Added

Monthly

Intensity response

Intensity gradation (4x4 & 2x16)

Image AddedImage Added

Monthly

Field uniformity

Field of rings

Image Added

Monthly

Field distortion

Field of rings

Image Added

Monthly

Stage drift during Z-stacking

3D crossing stairs

Image Added

Yearly

Stage drift during timelapse

Repositioning cross

Image Added

Yearly

Optical sectioning strength

Matrix of crosses

Image Added

Yearly

Accuracy of 3D reconstruction 

Sphere

Image Added

Yearly

Line spread function

Field of rings

Image Added

Yearly

Ring spread function

Field of rings

 

Image Added

Yearly

Method:

  • Step1: Image acquisition

Depending on what quality items to be checked with a certain fluorescence microscope, patterns can be imaged using the Argo SIM QC slide.

  • Step 2: Image analysis

Images of different pattens can be processed in the dedicated software Daybook 3.

Report:

Following the link below, we present the report on Quality Control (QC) in three imaging systems.