Microplates recommendations

Highly compatible plates for HTRF®

The right plate for a successful assay

Microplates are an important element of FRET signal detection. While most available microplate brands are compatible with HTRF technology and products, a number of parameters are critical to the quality of results.

  • FIA white plates from many suppliers remain the most appropriate standard. The signal to noise ratios obtained using white microplates do not differ greatly from those of black plates. Absolute counts, however, are significantly higher in white plates, thereby improving count statistics and subsequent assay reproducibility and sensitivity. NOTE: The use of white plates is mandatory for some assays and highly recommended when detection is performed using certain instruments, in particular on monochromator-based plate readers (Please refer to the package inserts).
  • Background levels may differ substantially among various plate types. Tissue-culture treated, or surface-treated plates may also be used.
  • Glass bottom plates produce suboptimal results and should not be used.

All the benefits of HTRF with ELISA plate ease of use

Taking these considerations into account, Cisbio’s 96w and 384w low volume microplates have been carefully designed to provide the best signal detection environment possible to all HTRF assays. They can be used in a no-transfer protocol with suspension cells or with a transfer step for adherent cells. These plates can be read on all HTRF certified readers, thus making their implementation in your experiments really easy

Provider Format Recommended volume Type Cat#
Cisbio 96-w low volume 20-30 µL White 66PL96001
Cisbio 384-w low volume 20-26 µL White 66PL384025

Other plates produced excellent results during our validation process for a broad range of HTRF assays. We have listed them in table. 1 below.

Provider Format Recommended volume Type Cat#
Corning 96-w half area TC treated  100 µL White 3688
Greiner 96-w half area TC treated  100 µL White 675083
Greiner 384-w 80-100 µL White 781075
Greiner 384-w low volume 15-20 µL White 784075
Greiner 384-w 80-100 µL White – TC* – sterile 781073
Greiner 384-w 80-100 µL White – TC* – sterile 781080
Greiner 384-w low volume 15-20 µL White – TC* – sterile 784080
Greiner 1536-w 5-10 µL White 783075
Greiner 1536-w 5-10 µL White – TC* – sterile 782080
Greiner 1536-w 5-10 µL White – TC* – sterile 782078
Greiner 1536-w 5-10 µL White, sterile 782073
Greiner 1536-w 5-10 µL White – TC* – sterile 782093
NUNC 384-w low volume 15-20 µL White, NT 264706

Table 1: a selection of recommended plates and corresponding volume per well

Whatever the plate type, assay volume should match well volume as closely as possible so that focalization of the reader excitation beam gives maximum energy delivery (Fig. 1) . For instance, when assay volumes are equal to or lower than 20 µl, it is better to use 384 low-volume or 1536-well plates rather than regular 384-well plates.

Well volume role in signal recovery
Figure 1: Signal recovery is highly dependent on assay volume/well volume ratio

Matching culture containers with signal detection formats

In case your experimental or culture conditions require you cultivate your cells in a specific flask or plate format prior to performing an assay, you can still benefit from an optimal reading by adapting the transfer step between culture format and 96w or 384w reading plate.

This Technical Note will help you performing that transfer step as smoothly as possible.