Presentation is loading. Please wait.

Presentation is loading. Please wait.

Relationship between A(OD) and %T Transmittance, T = P / P 0 % Transmittance, %T = 100 T Absorbance, A = log 10 P 0 / P A = log 10 1 / T A = log 10 100.

Similar presentations


Presentation on theme: "Relationship between A(OD) and %T Transmittance, T = P / P 0 % Transmittance, %T = 100 T Absorbance, A = log 10 P 0 / P A = log 10 1 / T A = log 10 100."— Presentation transcript:

1 Relationship between A(OD) and %T Transmittance, T = P / P 0 % Transmittance, %T = 100 T Absorbance, A = log 10 P 0 / P A = log 10 1 / T A = log 10 100 / %T A = 2 - log 10 %T

2 Beer Lamert’s Law

3 Reflection

4 Light scattering

5 reflection scattering For Solution: Scattering  1/ 4

6 UV-Vis Spectrum of Milk

7

8 Prism Diffraction grating

9 Spectrophotometer types -Single beam -Dual beam -Diode array

10 Single Beam - Spectrophotometer

11 Dual Beam - Spectrophotometer

12 Dual Beam – Single Detector

13 Diode Array - Spectrophotometer

14

15

16 NanoDrop

17 Bradford Assay

18

19

20 Substrate (S) and enzyme (E) combine to form the enzyme/substrate complex (ES). The complex then dissociates to yield enzyme (E) plus product (P).

21 Enzyme-Linked Immunosorbent Assay ELISA

22 LDH Cytotoxicity Assay

23 Endpoint vs Kinetic

24

25 Buffer Dilution V 1 x C 1 = Example: Need to make 1 L of 1mg/mL solution given 100mg/mL stock Example 2: Need to add component from 5.2x stock to 200mL of sample ?V 2 x C 2

26 Fluorescence is the emission of light by a substance that has absorbed light or other electromagnetic radiation of a different wavelength. George Gabriel Stokes named the phenomenon fluorescence in 1852. The name was derived from the mineral fluorite (calcium difluoride)

27

28

29 Molecular Orbital

30

31 Factors that influence on Fluorescence pH Solid state or Solution state Solvent

32 Vibrational and rotational relaxation AbsorbanceFluorescence Energy

33 The excitation and emission spectra of a fluorophore and the correlation between the excitation amplitude and the emission intensity. General diagram of the excitation and emission spectra for a fluorophore (left). The intensity of the emitted light (Em1 and Em2) is directly proportional to the energy required to excite a fluorophore at any excitation wavelength (Ex1 and Ex2, respectively; right).

34 The Stokes shift of the excitation and emission spectra of a fluorophore. Fluorophores with greater Stokes shifts (left) show clear distinction between excitation and emission light in a sample, while fluorophores with smaller Stokes shifts (right) exhibit greater background signal because of the smaller difference between excitation and emission wavelengths.

35

36 reflection Emission scattering Exitation

37

38 Emission Excitation Spectrofluorometer Detector monochromator

39 Emission Excitation Dichroic Mirror Microscope and Plate Reader Detector Filter

40 Optical Path Microplate Reader

41 http://www.chroma.com/products/catalog/11000_Series/11000v3 Filter and Dichroic Mirror

42 http://www.invitrogen.com/site/us/en/home/support/Research-Tools/Fluorescence-SpectraViewer.html

43 https://www.omegafilters.com/curvo2/index.php

44


Download ppt "Relationship between A(OD) and %T Transmittance, T = P / P 0 % Transmittance, %T = 100 T Absorbance, A = log 10 P 0 / P A = log 10 1 / T A = log 10 100."

Similar presentations


Ads by Google