Starna Fluorescence Reference Plates

Purpose

These references are designed for the routine qualification of fluorescence-based plate readers for fluorescence intensity and well-to-well reading reproducibility.

Description and Discussion

These references were formerly marketed by Matech.

96-well microplates containing inorganic fluorescent reference materials, available in both top- and bottom-reading configurations. Plate dimensions conform to international standards for biomedical screening.

The inorganic fluorophores used are inherently stable and do not suffer from photodecomposition or “bleaching”. Solvent evaporation is not an issue as these references are solid state. Four different fluorophores, identified by the wavelength of their principal emission peak, provide a range of excitation and emission wavelengths to match the user’s application:

Identification	PEAK EXCITATION WAVELENGTH (NM)	Excitation Range (nm)	Peak Emission Wavelength (nm)	Emission Range (nm)
Violet - 395	335	250 - 375	395	340 - 460
Blue - 455	325	250 - 410	455	410 - 550
Green - 517	280	250 - 490	530	480 - 600
Red - 615	280	250 - 530	525, 590, 615, 655, 705	470 – 715
Infrared - 800	440	380 - 680	702	620 - 820

Typical spectra

Note that spectra obtained on individual instruments will vary, especially when cut-off filters are used to distinguish between excitation and emission energies. Instrument resolution will also be a factor. The spectra above should be representative of what is achievable with modern plate reader instruments.

The Calibration Certificate supplied with each reference plate gives normalized emission spectrum data over the working range, plus relative intensity values for all the wells in the plate and the calculated coefficient of variation for those readings. Typical data (for a bottom reading BR615 plate) is shown below:

	1	2	3	4	5	6	7	8	9	10	11	12
A	0.941	0.957	0.953	0.957	0.952	0.954	0.95	0.949	0.927	0.925	0.918	0.913
B	0.952	0.951	0.953	0.96	0.956	0.957	0.946	0.932	0.938	0.908	0.918	0.931
C	0.956	0.957	0.96	0.96	0.951	0.949	0.946	0.948	0.929	0.926	0.918	0.916
D	0.951	0.962	0.961	0.965	0.961	0.952	0.952	0.945	0.941	0.938	0.928	0.921
E	0.965	0.962	0.96	0.966	0.96	0.961	0.957	0.957	0.945	0.953	0.931	0.939
F	0.972	0.969	0.948	0.966	0.963	0.958	0.963	0.958	0.942	0.935	0.932	0.965
G	0.987	0.968	0.964	0.958	0.965	0.967	0.97	0.982	0.971	0.945	0.963	0.96
H	1	0.971	0.967	0.971	0.977	0.983	0.982	0.973	0.944	0.942	0.938	0.947

CV: 1.83 %

The maximum coefficient of variation (CV) in fluorescence intensity across all wells is warranted to not exceed five percent.

Catalogue Number

Identification	Emission Peak Wavelengths (nm)	No. of wells	Top/Bottom Reading	CATALOGUE NUMBER
Violet 395	395	96	Top	TR-395
		96	Bottom	BR-395
Blue 455	455	96	Top	TR-455
		96	Bottom	BR-455
Green 517	530	96	Top	TR-517
		96	Bottom	BR-517
Red 615	525, 590, 615, 655, 705	96	Top	TR-615
		96	Bottom	BR-615
Infrared 800	702	96	Top	TR-800
		96	Bottom	BR-800

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Data sheet

Technique

Microplate References