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Installation Qualification (IQ) / Operational Qualification (OQ) Quality Control for Benchtop Microscopes 

Andor is proud to launch a revolutionary new initiative for light microscopy, developed through intensive collaboration with imaging experts in the scientific community. For years, microscopists have struggled with the challenge of quality control, and how to apply quantitative methods to research in the absence of fixed specifications and means of monitoring performance.

Andor's Installation Qualification (IQ) / Operational Qualification (OQ) programme, designed for its series of benchtop microscopes, changes that by promoting standardised procedures, reducing variability, and enhancing the reproducibility of results.

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Benefits of Andor’s IQ / OQ Programme

Quantitative Performance

All light microscopes represent an image which is defined by physical limitations. With this new approach to quantifiable performance, we can start to apply knowledge of the performance limitations, otherwise known as the information limit, to experiment design. This approach results in more tightly controlled experiments, allowing different methods and techniques.

Optimum Performance

Point-spread function (PSF) is a measure of system resolution, usually determined by imaging the spatial extent of a small point-source of light (a fluorescent microsphere for example), and then measured using an image analysis routine ((MetroloJ_QC plugin or PSFj).

Cédric Matthews and Fabrice P. Cordelieres: “MetroloJ: an ImageJ plugin to help monitor microscopes’ health,”

PSFj: know your fluorescence microscope

The graph on the right shows both lateral and axial resolutions of different types of fluorescent microscopes as a ratio of the theoretical full width half-maximum (FWHM).

Acceptable normalized resolution ratios, as determined by Faklaris et al, have a ratio value of 1.5 or less (green lines). Green emission channel (GFP) ratios presented.

Quality Control

Light microscopes are dynamic instruments, often used by a multitude of users with differing needs on a day-to-day basis. For researchers and lab owners who need to demonstrate system performance, our IQ / OQ programme provides the framework required to ensure your system is delivering the same performance on its first day as its last day.

Marker legend: WF = Widefield epifluorescence microscopes. SDCM = Spinning Disk Confocal Microscopes. LSCM = Laser Scanning Confocal Microscopes. BC43 = Andor Benchtop Confocal Microscopes.

Andor’s guaranteed minimum resolution ratios (1.47 lateral and 1.34 axial) when using a 60x 1.42 NA oil immersion objective lens are indicated by the orange lines. Example resolution performance measurements for several identical BC43 units using this type of objective lens are indicated by the orange triangle markers. Figure developed from data and specifications provided by Quarep.

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BC43 Microscope IQ / OQ Specification Table 

The specification table below details the imaging performance of BC43 microscopes equipped with a 60x 1.42 NA objective lens.

Our optional quality control service package will quantify the actual performance of your system within our published specifications. Every performance check carried out in your facility will result in a report provided to you detailing the actual measured performance. If any test fails during the scheduled periodic checks, our engineers will rectify to ensure continued performance within specification.

Channel Dependent Measurements

OQ Test Blue Channel Specification
405 nm Excitation
435-455 nm Emission
Green Channel Specification
488 nm Excitation
517-541 nm Emission)
Yellow Channel Specification
561 nm Excitation
580-610 nm Emission
Red Channel Specification
638 nm Excitation
671-745 nm Emission
Laser Power* P ≥ 12.5 mW P ≥ 8.5 mW P ≥ 9.5 mW P ≥ 12.0 mW
System Uniformity U ≥ 20% U ≥ 65% U ≥ 65% U ≥ 65%
Illumination Centering C ≥ 65% Centering performance for other channels exceeds our measuring sensitivity

* This test uses the epifluorescence (widefield) imaging mode of the microscope. All other tests utilize the confocal imaging mode of the microscope.

Channel-pair Measurements

OQ Test Blue-to-Green Chanel Pair Specification Green-to-Yellow Channel Pair Specification Green-to-Red Channel Pair Specification
Full Field of View Maximum Lateral Separation Distance (Lateral Co-registration) Max |rxy| (Full FOV) ≤ 480 nm Max |rxy| (Full FOV) ≤ 340 nm Max |rxy| (Full FOV) ≤ 620 nm
Full Field of View Average Axial Separation Distance (Axial Co-registration) Avg |rz| (Full FOV) ≤ 505 nm Avg |rz| (Full FOV) ≤ 410 nm Avg |rz| (Full FOV) ≤ 870 nm
Full Field of View Channel pair Co-registration (QUAREP Normalization) Max |rexp / rref| (Full FOV) ≤ 3.17 Max |rexp / rref| (Full FOV) ≤ 1.93 Max |rexp / rref| (Full FOV) ≤ 3.61
Central 30% Field of View Maximum Lateral Separation Distance (Lateral Co-registration) Max |rxy| (30% FOV) ≤ 180 nm Max |rxy| (30% FOV) ≤ 150 nm Max |rxy| (30% FOV) ≤ 205 nm
Central 30% Field of View Average Axial Separation Distance (Axial Co-registration) Avg |rz| (30% FOV) ≤ 505 nm Avg |rz| (30% FOV) ≤ 410 nm Avg |rz| (30% FOV) ≤ 870 nm
Central 30% Field of View Channel pair Co-registration (QUAREP Normalization) Max |rexp / rref| (30% FOV) ≤ 1.55 Max |rexp / rref| (30% FOV) ≤ 1.07 Max |rexp / rref| (30% FOV) ≤ 1.89

Single-channel Measurements

OQ Test Specification
Detector Intensity Response R2Int ≥ 0.96
Z-stage 3D Reconstruction Accuracy 0.97 ≤ G ≤ 1.03
XY-Stage Positioning and Repeatability Precision Max |Driftxy| ≤ 2 µm pk-pk
Central 30% Field of View Average Lateral Resolution Avg FWHMxy (30% FOV) ≤ 280 nm
Central 30% Field of View Average Lateral Resolution (QUAREP Normalization) Avg FWHMxy / resox/y (30% FOV) ≤ 1.47
Full Field of View Average Lateral Resolution Avg FWHMxy (Full FOV) ≤ 280 nm
Full Field of View Avearage Lateral Resolution (QUAREP Normalization) Avg FWHMxy / resox/y (Full FOV) ≤ 1.47
Central 30% Field of View Average Axial Resolution Avg FWHMz (30% FOV) ≤ 725 nm
Central 30% Field of View Average Axial Resolution (QUAREP Normalization) Avg FWHMz / resoz (30% FOV) ≤ 1.34
Full Field of View Average Axial Resolution Avg FWHMz (Full FOV) ≤ 725 nm
Full Field of View Average Axial Resolution (QUAREP Normalization) Avg FWHMz / resoz (Full FOV) ≤ 1.34
System Vibration * σxy ≤ 40 nm
Contamination and Background Artifacts Transmitted light Brightfield and Fluorescence Background images must match factory reference images
(See test description for more details)

* This test uses the epifluorescence (widefield) imaging mode of the microscope. All other tests utilize the confocal imaging mode of the microscope.

Why is Quality Control Important?

Understanding the exact performance of a microscope is crucial for experiment validation. Our goal is not only to provide owners and laboratories with a quality control programme to ensure consistent system performance, but also to provide researchers with the information they need to plan their experiments.

Our system tests provide researchers with hard data on the individual system parameters that influence their experiments. The tests use a mix of samples and software best adopted to each parameter.

The programme has been developed in collaboration with QUAREP-LiMi, an initiative with over 600 members from academia and industry, which is aiming to improve the reproducibility of light microscopy experiments in life and material sciences.

In this video, Dr. Glyn Nelson outlines how this approach to quality control contributes to the credibility of research conducted with microscopy techniques, and advances the pursuit of scientific knowledge.