Plate reader for volume verification and recommended assays?

Plate Readers
1

Hello everyone,

I am looking for advice on what kind of plate-reader we would benefit most from and what system is generally preferred from a user perspective.

Today, we are mostly running absorbance based assays using tartrazine dyed reagents (we produce our own reagents) to verify pipetting performance. We are measuring in the volume span of 0.1µL to 50µL.

I know there are a lot of different method much more likely to show us the true pipetting performance - would anyone have a specific method to recommend? Quant-iT kits or qubit seems promising. Any advice on designing a dual-dye assay?

Right now Varioskan lux looks like an alternative as it might cover some functions requested by other departments, main drawback is that it is rather large and bulky. Does anyone have experience with this system?

Best regards,
Wictor

Edit: Are there any plate reader that can do the fluorescence and absorbance measurement for each well subsequently? I.e., for each well do both measurements and then proceed to next well.

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I have a SpectaMax M5 in my lab and it does the job well enough. I found a linear range for tartrazine between 1-7% (w/v) final concentration in 100 uL sample well (standard 96PCR plate), with OD between 0.5-2.5 before the R2 began to break down. I ran all my volume verification with on-plate std curve, but also found very little variation in the regression over 20 different plates so an external curve might have been acceptable. This method and well volume would be specific enough for your volume range down to ~0.5 uL unless you reduced the well volume and confirmed the linear range at that lower well volume. (or a better read plate with higher RFU, or better plates like A/2 chimney well, etc…)

The M5 is bulky, and would need multiple “experiments” set up within the same protocol to read Flu and Abs, but you’d need to switch between top-read and bottom read regardless to get better OD, which requires an adapter on the M5. Not using the adapter would risk tightening the linear range and subsequently the volume range you could measure - at least with the M5.

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