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 | Monthly | |
Intensity response | Intensity gradation (4x4 & 2x16) | Monthly | |
Field uniformity | Field of rings | Monthly | |
Field distortion | Field of rings | Monthly | |
Stage drift during Z-stacking | 3D crossing stairs | Yearly | |
Stage drift during timelapse | Repositioning cross | Yearly | |
Optical sectioning strength | Matrix of crosses | Yearly | |
Accuracy of 3D reconstruction | Sphere | Yearly | |
Line spread function | Field of rings | Yearly | |
Ring spread function | Field of rings |
| 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.
Olympus BX53 Fluorescent Microscope
Olympus VS120 Slide Scanner
DeltaVision Elite Deconvolution Microscope
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