EPOCH Microplate Spectrophotometers 2012.Sep. BioTeks product - - PowerPoint PPT Presentation

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EPOCH Microplate Spectrophotometers 2012.Sep. BioTeks product - - PowerPoint PPT Presentation

Aug 14, 2022 445 likes •630 views

EPOCH Microplate Spectrophotometers 2012.Sep. BioTeks product range: microplate instrumentation Washers Dispensers Pipetors Automation Absorbance Fluorescence Luminescence Multi-detection Monochromator Optical System Data

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SLIDE 1

EPOCH

Microplate Spectrophotometers

謝嚴蔚 2012.Sep.

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SLIDE 2

BioTek’s product range: microplate instrumentation

Washers Dispensers Pipetors Automation Absorbance Fluorescence Luminescence Multi-detection

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SLIDE 3

Xenon Flash Lamp Order Sorting Filters Diffraction Grating

Lens Photodiode Microplate

Data

Monochromator Optical System

Optical fibers

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SLIDE 4

100 250 500 750 1100

Xenon Flash Lamp

  • Continuous spectrum – 200~999 nm
  • Long life – 1 billion flashes
  • No warm-up required

Key Specifications – UV-Visble Spectrophotometer

Monochromator

  • No filters
  • Continuous wavelength selection
  • Spectral scans

Single channel optics Detector- Photodiode

Relative Energy Wavelength 100 250 500 750 1100

Xe Tg

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SLIDE 5

Enhanced Reading Mode

Plate

8 Flashes OD Measured 0.923

If less than 2.0 OD then on to next sample

8 Flashes OD Measured 2.700

If Greater than 2.0 OD then 64 more flashes, then on to next sample

64 Flashes OD Measured 2.678 8 Flashes OD Measured 0.675 8 Flashes OD Measured 1.234

WHY ?

ERROR 1 OPTICAL DENSITY 2 3 4

± 1% ± 0.005 NON ENHANCED ENHANCED

Ensures the highest quality readings

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SLIDE 6

Microplates (model dependent)

  • Microplates up to 20.3 mm in height

– 6-well – 12-well – 24-well – 48-well – 96-well – 384-well

  • Terasaki trays

– 60-well – 72-well – 96-well

  • Up to 8 Bio-Cells for 1 cm readings
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SLIDE 7

Utilizes Bio-cell 1.0 cm Quartz Cuvette for Vertical Photometery 1cm Adapter Plate for Bio-Tek Microplate readers

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SLIDE 8

Major Applications

  • Molecular Biology
  • Protein Quantitation
  • ELISA
  • Cell Viability/Cytotoxicity
  • Miscellaneous
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SLIDE 9

Molecular Biology

  • Nucleic Acid Quantitation
  • DNA
  • RNA
  • Oligonucleotides
  • Nucleic Acid Quality Control
  • 260/280 Ratio (protein)
  • 260/240 (EDTA)
  • 260/270 (phenol)
  • 320 (particulates)

** UV Plate

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SLIDE 10

Nucleic Acid Quantitation

  • Direct Quantitation : 280 nm (DNA Concentration = OD x 50)
  • Colorimetric Assays
  • Burton Method
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SLIDE 11

Pathlength Correction

Horizontal Photometry Light Source Detector Absorbing solution Vessel

? cm

  • Beer-Lambert Equation:

O.D. = (ε) x (Concentration) x (pathlength)

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SLIDE 12

Protein Quantitation

  • 1. Direct quantitation: 280 nm
  • 2. Colorimetric：

– Lowry method: 660 nm – Bradford method: 595 nm – BCA method: 562 nm

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SLIDE 13

ELISA

Antibody Enzyme labelled antibody Sample antigen

w a s h

1

w a s h

1 3

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SLIDE 14

ELISA

  • Horseradish peroxidase
  • OPD 450 nm & 492 nm
  • TMB 650 nm & 450 nm
  • ABTS 414 nm
  • Alkaline phosphatase
  • pnpp 405 nm
  • β-Galactosidase
  • ONPG 420 nm
  • Urease
  • urea bromocresol 588 nm
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SLIDE 15

Cell-based assays

Cell viability/cytotoxicity

  • MTT

570 nm

  • XTT

492 nm

  • WST-1

450 nm

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SLIDE 16

Spectral scanning

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SLIDE 17

Software Control

  • All control via USB port
  • Gen5 software

– Operating system of choice – Included with instrument – Increased flexibility – Robotic applications

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SLIDE 18

Any Questions?