Tecan Infinite 200 Pro

Multifunctional Microplate Reader with
IInfinite fi it 200® PRO
injector option

Measurement techniques supported by the
Infinite M200 Pro:
Absorbance:
‐ Single point and scanning
‐ Single point and scanning
with injectors
Fluorescence:
‐ Fluorescence Intensity top and bottom
‐ Time‐Resolved Time Resolved Fluorescence
‐ Fluorescence Resonance Energy Transfer
‐ Flash Fluorescence (with injectors)
TECAN
IInfinite fi it
M200 Pro
Luminescence:
‐ Gl Glow LLuminescence i
‐ Flash Luminescence
‐ Bioluminescence Resonance Energy Transfer
Absorbance and
Fluorescence:
NNanoquant: t
Small volume measurements,
2 ul sample

Technical specifications p Infinite M200 Pro
General specifications:
� Microplates to be measured: from 66‐well well to 384‐well 384 well plates
� Temperature Control: from 5 °C above ambient up to 42 °C
� Plate Shaking: Linear and orbital shaking, amplitude from 1 –6 mm in 0.5 mm steps
� Light Source: High energy xenon flash lamp
� Detectors: Fluorescence ‐> PMT; Luminescence ‐> Low dark count PMT photon‐
counting electronics; Absorbance ‐> Silicon photodiode
Ab Absorbance b specifications: ifi ti
� Wavelength range: 230 – 1000 nm
� Accuracy: < ±(1 % + 10 mOD); ≤ ±1.5 nm λ > 315 nm; ≤ ±0.8 nm λ ≤ 315 nm;
Fluorescence specifications:
� Wavelength Range: Excitation: 230 – 850 nm; Emission: 280 – 850 nm
� Detection limit (fluorescein, 96 well plate, 200 μl): < 20 pM (top); 100 pM (bottom)
� Europium detection limit (96 well plate, 200 μl ): < 5 pM (typical value)
� Reproducability (at 25nM fluorescein) < 2%

Technical specifications p Infinite M200 Pro
Luminescence specifications:
�� Wavelength Range: 380 – 600 nm
� Linear dynamic range: at least 6 orders of magnitude
� Integration time per well: 100 – 20,000 ms
�� Cross Talk (black plate): 00.01% 01%
� Attenuation of light: 100 (OD2 attenuation filter), 1 (no attenuation)
� ATP detection limit (96 well plate, 200 μl volume, 1000 ms integration time):
3 fmol/well (glow); 80 amol/well (flash)
Injector specifications:
� Accuracy: < 10% for injection volume of 10 μl; < 2% for injection volume of 100 μl;
< 0.7% for injection volume of 450 μl
� Precision: < 10% for injection volume of 10 μl; < 2% for injection volume of 100 μl;
< 0.7% for injection volume of 450 μl

Comparison measurements on Tecan multiplate
reader vs Thermo “Nanodrop”
Measurements of DNA at different known concentrations, 2 μl volumes:
TECAN multiplate reader
Thermo Nanodrop
Known
CConcentration i
[ng/ml]
Concentration
measured d by b
NanoDrop [ng/ml]
Concentration
measured d by b
Tecan [ng/ml]
0.05 0.84 0
0.5 3.85 0.1
5 8.61 5.4
50 48.71 52.2
500 443.83 492.7
Note: with the TECAN Nanoquant, 16 samples can be measured in parallel vs 1 on Nanodrop.
Note: fluorescence and absorbance can be measured on TECAN Nanoquant (e.g. for labelling efficiency)
vs absorbance only on Nanodrop.

Typical Readout for ELISA (absorbance)
Absorbance measurement at 405nm with a reference wavelength of 650nm
Calculated difference between measurement and reference measurement
Mycoplasma species (Sample) Neg. control Pos. control
M. arginini 0.1747 0.1334 2.7127
M. hyorhinis 0.22325 0.152 1.2394
A. laidlawii 0.1817 0.1063 1.3544
M. orale 0.16455 0.145 2.6902
Sample

Typical Readout for luminescence
Luminescence readings for a Mycoplasma detection test based on a bioluminescent
reaction: Reading A before substrate addition, Reading B after substrate addition
� Transparent plate reading
Reading A Reading B Ratio (B/A)
Pos. control 537 22624 42.13035382
Neg. control 618 132 0.213592233
bEND3 (Sample) 311 144 0.463022508
The bEND3 brain endothelial cells are negative for Mycoplasmal enzymes.
� Black plate reading
Reading A Reading B Ration (B/A)
Pos. control 42 2767 65.88095238
Neg. control 29 ‐1 ‐0.034482759
Both transparent and black plates can be adequately used for measurements. Black plates have
lower overall signal, but better signal to background ratio.

Typical Readout for fluorescence
Standard curve for fluorescence reading of different known concentrations of
fluorescein sodium salt
� Excitation: 485 nm
� Emission: 535 nm
Fluorescennce
intensity
45000
40000
35000
30000
25000
20000
15000
10000
5000
0
1 5 50 100 250 500 1000
Fluorescein concentration (µM)

Summary
The TECAN Infinite 200 Pro at the BNL lab offers:
� High accuracy multiplate readings for fluorescence,
absorbance and/or luminescence
� Temperature control, dual injectors, shaking
� Software with a user friendly interface
� Nanoquant option for 2 μl sample volumes
(absorbance and/or fluorescence), equal or
superior accuracy to “ “Nanodrop” d ” instrument.
Link to recharge information:
http://www.cnsi.ucsb.edu/facilities/nanostructures/biological/recharge‐rates.pdf