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TABLE OF CONTENTS+ _9 D6 F" {+ H
• Chapter 1 Flow Cytometry Instrumentation6 c2 G* {: r2 G* G
• Introduction
; x1 @0 K6 b6 A• Unit 1.1 Overview of Flow Cytometry Instrumentation
. E y0 H( C+ l' g q• Unit 1.2 Fluidics
8 R- ^/ O& a6 P3 a/ s• Unit 1.3 Standardization, Calibration, and Control in Flow Cytometry" f& h1 |5 ^* @
• Unit 1.4 Establishing and Maintaining System Linearity
9 \, ~$ \1 H; X# _• Unit 1.5 Optical Filter Sets for Multiparameter Flow Cytometry. m( {+ U* R) A: y1 v
• Unit 1.6 Laser Beam Shaping and Spot Size
1 s2 C" Y0 ?( T2 t! |• Unit 1.7 High‐Speed Cell Sorting- J- s U/ J7 t* W1 f1 T
• Unit 1.8 Principles of Gating
$ `) k: {* U. R1 M- a. s• Unit 1.9 Lasers for Flow Cytometry
+ x' P$ ]+ L0 k' ~7 L5 ?• Unit 1.10 Techniques for Flow Cytometer Alignment
) Q) O3 C' s% I. y• Unit 1.11 Flow Cytometers for Characterization of Microorganisms
( u% n; D$ d. V• Unit 1.12 Principles of Resonance Energy Transfer
- t& H1 H+ i- ~3 h( h• Unit 1.13 Light Scatter: Detection and Usage. @/ s: C0 h6 G% w
• Unit 1.14 Compensation in Flow Cytometry
0 W; I# A, u* v• Unit 1.15 Time‐Resolved Fluorescence Measurements
' {) }' ]! g% E6 |• Unit 1.16 Simultaneous Analysis of the Cyan, Green, and Yellow Fluorescent Proteins7 e y0 X) q" H6 Z4 p
• Unit 1.17 Plug Flow Cytometry! [$ a1 ]6 d6 V$ H8 P
• Unit 1.18 Dynamic Thermoregulation of the Sample in Flow Cytometry# R4 i' z! k: h% S
• Unit 1.19 Excitation and Emission Spectra of Common Dyes
V) O0 {9 A) p• Unit 1.20 Characterization of Flow Cytometer Instrument Sensitivity5 v2 n5 ^' b, S) d, U% g
• Unit 1.21 Separation Index: An Easy‐to‐Use Metric for Evaluation of Different Configurations on
0 }6 T/ s( a3 S# { K. @the Same Flow Cytometer
( C Y, y% R8 ~% d% [• Unit 1.22 Fundamentals of Acoustic Cytometry
% r2 m; I& n6 v5 c( }. y) i• Unit 1.23 Pulse Width for Particle Sizing
( I p: a. w0 M* n7 b9 s% h7 X• Unit 1.24 Practical Issues in High‐Speed Cell Sorting2 y% Q- I* f/ j6 w! K$ H5 I
• Unit 1.25 Capture of Fluorescence Decay Times by Flow Cytometry! U* ]1 g* q- h/ B8 s7 \
• Unit 1.26 Fountain Flow Cytometry& T3 S: |. K0 X
• Unit 1.27 Spectral Flow Cytometry
9 O1 d0 o( Q) Y8 F• Unit 1.28 Evaluation and Purchase of an Analytical Flow Cytometer: Some of the Numerous
' D M+ p' M( S Y" }1 T- ^! eFactors to Consider' s7 n: {& |9 q1 g
• Unit 1.29 Quantitative Flow Cytometry Measurements in Antibodies Bound per Cell Based on a
: X+ @$ C0 F" |4 JCD4 Reference
& j$ q5 I- k4 C9 X/ [& f• Unit 1.30 A Quantitative Method for Comparing the Brightness of Antibody‐dye Reagents and
2 ~6 W4 \7 A% G2 P$ fEstimating Antibodies Bound per Cell k) W! a6 U1 ~2 q |# y; _
• Chapter 2 Image Cytometry Instrumentation
* b9 C- C0 w7 P0 b1 X1 M: d' h% t: H* A• Introduction) x5 P: Y, U% p* Z: Y# U
• Unit 2.1 Contrast Enhancement in Light Microscopy
3 G6 Q' J9 Q% ]' [• Unit 2.2 Microscope Objectives
' d1 W2 \/ l! z( R5 P7 }1 n9 ~% O/ p• Unit 2.3 Light Microscopy Digital Imaging) ^& [1 H) i7 q% n- V+ t, o7 W1 R
• Unit 2.4 Optical Filters for Wavelength Selection in Fluorescence Instrumentation
, L l+ X; S c3 D( B6 ~4 v• Unit 2.5 Digital Fluorescence Microscopy
2 D# l. P D h/ a• Unit 2.6 Calibration: Sampling Density and Spatial Resolution
; d! w h4 z! m/ r. ]$ N' g( x• Unit 2.7 Microscope Alignment
) @ E8 ~$ V6 q• Unit 2.8 Confocal Microscopy: Principles and Practices
+ p* L& J0 g a( U: Y) @• Unit 2.9 Multi‐Photon Imaging
9 x# X/ ]) m! \* z" Z• Unit 2.10 Scanning Laser Cytometry0 X/ u6 |3 w8 Y; _" `
• Unit 2.11 Shading Correction: Compensation for Illumination and Sensor Inhomogeneities
$ E7 W7 a. p2 e4 E1 `7 t• Unit 2.12 Photobleaching Measurements of Diffusion in Cell Membranes and Aqueous Cell
2 a. ^ L2 r. b/ x& G2 ]! ]Compartments; q. ~; u& y9 T# b8 S) v
• Unit 2.13 Optimizing Laser Source Operation for Confocal and Multiphoton Laser Scanning; A8 _% {# j, }' f+ B
Microscopy2 f; b" T7 |; W1 x
• Unit 2.14 Methods and Applications of Laser‐Enabled Analysis and Processing (LEAP)+ C! A8 k! A7 F; K. {
• Unit 2.15 Measurement of Molecular Mobility with Fluorescence Correlation Spectroscopy4 X1 l& l% `8 ~; v& F
• Unit 2.16 Evaluation and Purchase of Confocal Microscopes: Numerous Factors to Consider$ N f- t& o+ p9 j
• Unit 2.17 Super‐Resolution Microscopy: A Comparative Treatment5 {8 x4 ~! q; r, X6 o8 I
• Unit 2.18 Quantitative Fluorescent Speckle Microscopy (QFSM) to Measure Actin Dynamics/ z7 R9 N1 Z" ?0 w4 X5 R6 {
• Unit 2.19 Analysis of Protein and Lipid Dynamics Using Confocal Fluorescence Recovery After
5 F4 n; T" a `' q. W( G. APhotobleaching (FRAP)
. D$ P" C, W; V4 l# w5 W• Unit 2.20 Comparative and Practical Aspects of Localization‐Based Super‐Resolution Imaging
# G/ q9 t9 ?2 `• Unit 2.21 Building a Live Cell Microscope: What You Need and How to Do It
& W k& z. {9 l" G8 q• Unit 2.22 How to Build a Time‐Gated Luminescence Microscope; T/ m p5 j6 e; x& j# x7 {: @
• Unit 2.23 FRET Imaging by Laser Scanning Cytometry on Large Populations of Adherent Cells
$ X4 F' s- `0 ?) s' t• Chapter 3 Safety Procedures and Quality Control
8 X; d' R" W+ e Z* @9 y k8 R• Introduction
7 |5 W; m2 [" ~8 F) F5 P• Unit 3.1 Principles of Quality Control2 U0 I2 X4 H, j/ z
• Unit 3.2 Components of Quality Control
5 _8 b' V8 y8 O- s! z• Unit 3.3 Testing the Efficiency of Aerosol Containment During Cell Sorting, r# J4 \' L: P5 g
• Unit 3.4 Safe Use of Hazardous Chemicals' H; j' r. m2 W+ L* P
• Unit 3.5 Method for Visualizing Aerosol Contamination in Flow Sorters" {8 H9 Q/ Z2 s2 a a7 C! o' p7 Z
• Unit 3.6 Standard Safety Practices for Sorting of Unfixed Cells. Q6 i- M0 q8 n& {
• Chapter 4 Molecular and Cellular Probes
1 F1 o* t8 r7 m• Introduction
( g$ h, R7 J- x1 ]• Unit 4.1 Titering Antibodies6 O1 X% R# E% b8 O( f$ j: s
• Unit 4.2 Conjugation of Fluorochromes to Monoclonal Antibodies
. j" j( M" `! ?) I• Unit 4.3 Nucleic Acid Probes
: }8 M8 z% ~7 F9 l• Unit 4.4 Cellular Function Probes* r- h0 E; ~" x! N6 i; V! H% O1 x+ m
• Unit 4.5 Spectroscopic Analysis Using DNA and RNA Fluorescent Probes1 c, E" c% c( i
• Unit 4.6 Flow Cytometric Sorting of Bacterial Surface‐Displayed Libraries7 x7 m& f; D# S0 h) X* V1 V+ d
• Unit 4.7 Construction and Screening of Antigen Targeted Immune Yeast Surface Display
- |% D8 L% B" ^1 f9 J4 ?Antibody Libraries
/ l; V: j8 m6 i4 u j* |• Chapter 5 Specimen Handling, Storage, and Preparation
. @$ [$ {4 d5 ?$ j3 T% x• Introduction
" v( A% X+ E, [* E4 U2 J• Unit 5.1 Collection, Storage, and Preparation of Human Blood Cells4 y# ]% ~7 P) ?
• Unit 5.2 Handling, Storage, and Preparation of Human Tissues+ f$ A1 R. ~- u% I3 @& J
• Unit 5.3 Flow Analysis and Sorting of Plant Chromosomes
" U, j6 p3 q' H: J• Chapter 6 Phenotypic Analysis# t8 s" e& t: _; k" G% K. L
• Introduction* C6 t, F6 \# D
• Unit 6.1 Quality Control in Phenotypic Analysis by Flow Cytometry
# W/ b4 u& ]# m3 p! {• Unit 6.2 Immunophenotyping1 O* u8 @: \, Z" M# W! O* {
• Unit 6.3 High‐Sensitivity Immunofluorescence/Flow Cytometry: Detection of Cytokine2 S$ R7 E8 R7 {3 P6 |
Receptors and Other Low‐Abundance Membrane Molecules! X3 P+ U( w( u( a7 H0 `" V" M2 {/ k. F
• Unit 6.4 Enumeration of CD34+ Hematopoietic Stem and Progenitor Cells2 E% X* z$ J+ D! _* ]1 K4 m1 u
• Unit 6.5 Immunophenotypic Analysis of Peripheral Blood Lymphocytes' k* o2 g- }+ G9 w* n$ g2 n& z
• Unit 6.6 Immunophenotypic Analysis of Human Mast Cells by Flow Cytometry1 X$ j% E! j* l1 c, V2 P, d/ G2 Q
• Unit 6.7 Measurement of CD40 Ligand (CD154) Expression on Resting and In Vitro–Activated T
$ y* c( [8 D2 s4 E/ r( v$ G1 P& g5 UCells h, ~: b0 p1 u* Y4 h$ W9 y( O
• Unit 6.8 Enumeration of Absolute Cell Counts Using Immunophenotypic Techniques: Q. ^7 V9 r# t( Q4 w/ l9 b
• Unit 6.9 Immunophenotypic Identification, Enumeration, and Characterization of Human" P/ h! ]/ p" a' H
Peripheral Blood Dendritic Cells and Dendritic‐Cell Precursors
' A. g5 V0 o8 s) J2 a9 m• Unit 6.10 Immunophenotypic Analysis of Platelets
w+ Z1 c# u2 T) _$ l/ q• Unit 6.11 Immunophenotypic Analysis of PNH Cells3 R) F0 r" D$ U* E& _/ f3 Q
• Unit 6.12 Quantitative Flow Cytometric Analysis of Membrane Antigen Expression( d3 g! t1 ]8 k/ g
• Unit 6.13 Immunophenotyping Using a Laser Scanning Cytometer) f3 b6 x0 m+ ?' d" n
• Unit 6.14 Enzymatic Amplification Staining for Cell Surface Antigens: N r* G) T9 k2 B u! P
• Unit 6.15 Whole Blood Analysis of Leukocyte‐Platelet Aggregates
3 x4 C" y0 T5 x( f% {' J- i! E• Unit 6.16 Flow Cytometric Assessment of HLA Alloantibodies. d6 N/ R- O( c, z$ u3 J% I
• Unit 6.17 Enumeration of Fetal Red Blood Cells, F Cells, and F Reticulocytes in Human Blood! E; k$ q8 U q4 U9 i* x4 Y
• Unit 6.18 Identification of Human Antigen‐Specific T Cells Using MHC Class I and Class II, d" p. e8 y1 I" R# P
Tetramers
& s, @1 z5 @$ z, J, q* Z• Unit 6.19 ZAP‐70 Staining in Chronic Lymphocytic Leukemia1 F9 b7 `/ U& T0 @
• Unit 6.20 Multiparameter Analysis of Intracellular Phosphoepitopes in Immunophenotyped Cell
! O: ]+ @. P4 Z+ X: [; MPopulations by Flow Cytometry
& N& d. m8 L7 \8 `6 g8 B• Unit 6.21 Ten‐Color Immunophenotyping of Hematopoietic Cells
0 ~' `; H8 ^3 }1 m6 E* n• Unit 6.22 Flow Cytometric Screening for the HLA‐B27 Antigen on Peripheral Blood Lymphocytes
0 Z% p+ V5 Z7 J" ~3 N• Unit 6.23 Immunophenotyping of Plasma Cells
! Y! H6 b* n2 M9 D• Unit 6.24 Flow Rate Calibration for Absolute Cell Counting Rationale and Design o: Z8 l8 U n+ h) m; B
• Unit 6.25 Flow Cytometric Immunophenotyping of Cerebrospinal Fluid
9 A1 J, Q* M \• Unit 6.26 Calibration of Flow Cytometry for Quantitative Quantum Dot Measurements
: O1 l' z% M. A( |• Unit 6.27 Assessment of Beta Cell Viability5 V ^7 L8 I! S9 N/ A/ i5 n
• Unit 6.28 Measurement of T Cell Activation After 16‐hr In Vitro Stimulation with Concanavalin A
( U S6 i: s/ ~( y• Unit 6.29 Titration of Fluorochrome‐Conjugated Antibodies for Labeling Cell Surface Markers on
! n; w3 U8 Q, N% F* S! ]' J+ NLive Cells; s& o6 v* [( v' R% S
• Unit 6.30 Phenotypic Analysis Using Very Small Volumes of Blood% c7 W1 C/ C; ]
• Unit 6.31 Fluorescent Cell Barcoding for Multiplex Flow Cytometry+ V7 b8 b6 M$ G: j; V5 |: n
• Unit 6.32 Quantitative Assessment of Pancreatic Islets Using Laser Scanning Cytometry4 ~7 u8 z% t7 B! N h9 k# L2 @2 \
• Unit 6.33 Three‐Dimensional Second‐Harmonic Generation Imaging of Fibrillar Collagen in
& ^+ }- L( F$ w' XBiological Tissues) K- f$ Z$ L* Q- M! w; } e
• Unit 6.34 Flow Cytometry–Based Cytotoxicity and Antibody Binding Assay
2 {2 _6 h% U( X1 Q8 q1 s• Unit 6.35 Quantification of Th1 and Th17 Cells with Intracellular Staining Following6 _5 b* x, n3 Z1 a( Z& w7 ]
PMA/Ionomycin Stimulation
5 Y% S% g( B4 e$ r) x7 g: ^/ `+ h• Unit 6.36 Whole Blood Measurement of Histone Modifications Linked to the Epigenetic1 z" ~2 L5 T# R4 b
Regulation of Gene Expression) z. Z( ^2 y; h: J" [" D
• Unit 6.37 High‐Sensitivity Detection of PNH Red Blood Cells, Red Cell Precursors, and White8 p1 ^- P- d8 |5 Q; k( q
Blood Cells, w. \, I: h+ q. T+ P
• Unit 6.38 Method for DNA Ploidy Analysis Along with Immunophenotyping for Rare Populations
/ m: Q4 b2 T8 S zin a Sample using FxCycle Violet
' s9 }& }2 }8 ]1 }• Chapter 7 Nucleic Acid Analysis
9 x& }" G/ E+ A) T• Introduction
/ } i T3 Y7 l; T2 {: @• Unit 7.1 Overview of Nucleic Acid Analysis8 q% N* J' a' j$ Q) P$ c
• Unit 7.2 Critical Aspects in Analysis of Cellular DNA Content( S2 n" k2 |- I+ w7 J( o* s
• Unit 7.3 Differential Staining of DNA and RNA% _9 |( C5 n2 I1 z& o
• Unit 7.4 Analysis of DNA Content and DNA Strand Breaks for Detection of Apoptotic Cells
/ m+ ?& M+ W6 y• Unit 7.5 DNA Content Measurement for DNA Ploidy and Cell Cycle Analysis" R9 r! ^* y3 ~
• Unit 7.6 Analysis of Nuclear DNA Content and Ploidy in Higher Plants
+ B' B+ d( w2 ]6 Q* k( N( y' r$ k• Unit 7.7 Analysis of DNA Content and BrdU Incorporation
( V1 H1 W9 L2 i: [• Unit 7.8 Analysis of DNA Denaturation# \' U2 a1 |- ~4 [5 O2 `
• Unit 7.9 Bivariate Analysis of DNA Content and Expression of Cyclin Proteins
' [: |- m$ |- r• Unit 7.10 Flow Cytometric Analysis of Reticulated Platelets' I9 i. v! b* [
• Unit 7.11 Assessment of Viability, Immunofluorescence, and DNA Content. A5 j& `1 ]: S6 @& m8 n! n# K
• Unit 7.12 Flow Cytometric Analysis of RNA Synthesis by Detection of Bromouridine, @# y# N9 U" x8 l9 X4 j
Incorporation
- h5 N) t# j9 O# W• Unit 7.13 Sperm Chromatin Structure Assay for Fertility Assessment* v# x" w- s% Y
• Unit 7.14 Analysis of Cell Proliferation and Cell Survival by Continuous BrdU Labeling and
- c6 E2 \( n' m: ]2 zMultivariate Flow Cytometry+ j9 L" Q3 \5 F/ D5 h
• Unit 7.15 Ultraviolet‐Induced Detection of Halogenated Pyrimidines (UVID)
# {- P0 L/ m1 I4 Y5 D• Unit 7.16 Analysis of DNA Content and Green Fluorescent Protein Expression8 J* X7 J/ x1 B4 D) {' Q! c
• Unit 7.17 Analysis of Viral Infection and Viral and Cellular DNA and Proteins by Flow Cytometry7 c; _0 R! |8 |1 E# m
• Unit 7.18 Apoptosis Signaling Pathways
0 a6 x: d! `- _" S" w2 B! X8 H$ G• Unit 7.19 Flow Cytometry of Apoptosis/ A7 Q5 v4 i5 Y" h2 S, ^
• Unit 7.20 Analysis of Fine‐Needle Aspirate Biopsies from Solid Tumors by Laser Scanning! n% {" C: E a9 L& |8 y
Cytometry (LSC)+ T% @! h6 B% D6 l
• Unit 7.21 Measurement of Cytogenetic Damage in Rodent Blood with a Single‐Laser Flow
, f) r& l8 L- L" UCytometer
8 Y% F; K' p- ~• Unit 7.22 Analysis of Tissue Imprints by Scanning Laser Cytometry. c9 M9 X9 p z0 h: V# E
• Unit 7.23 Cell Cycle Analysis of Budding Yeast Using SYTOX Green# m/ K+ a [8 W1 J/ H2 j
• Unit 7.24 Detection of Mitotic Cells
! X' r' Q% o! P2 u• Unit 7.25 DRAQ5 Labeling of Nuclear DNA in Live and Fixed Cells, J0 `5 \& J% [) h0 |) r) k4 `/ s
• Unit 7.26 Assessment of Telomere Length, Phenotype, and DNA Content
# q( O4 g5 l# G0 y2 V( P7 k2 |• Unit 7.27 Detection of Histone H2AX Phosphorylation on Ser‐139 as an Indicator of DNA
* X: P. ^1 ?8 S# \9 bDamage (DNA Double‐Strand Breaks)) J! w( n( h9 j+ y U
• Unit 7.28 RNA and DNA Aptamers in Cytomics Analysis( `) F0 _* i% D# i
• Unit 7.29 Nuclear DNA Content Analysis of Plant Seeds by Flow Cytometry/ [! m5 X8 y% j. }
• Unit 7.30 Estimation of Relative Nuclear DNA Content in Dehydrated Plant Tissues by Flow, ]! {7 |- ]! ~, X- Y6 l8 r% \
Cytometry
: s! p1 }! j" j0 A1 H2 W• Unit 7.31 Assessment of Cell Proliferation by 5‐Bromodeoxyuridine (BrdU) Labeling for% _# C6 o/ w# q& \, ]/ `! m1 |, V
Multicolor Flow Cytometry
3 X7 q" p" |) H• Unit 7.32 Uncompensated Polychromatic Analysis of Mitochondrial Membrane Potential Using
0 x- Q4 B$ v3 s8 t& M% n- Z, `9 XJC‐1 and Multilaser Excitation3 M5 _6 Q2 b, T- j
• Unit 7.33 SYTO Probes: Markers of Apoptotic Cell Demise
/ B3 T/ P5 o0 D- S• Unit 7.34 Cell Proliferation Method: Click Chemistry Based on BrdU Coupling for Multiplex; U8 e+ G7 n, ]' x: X o
Antibody Staining
2 ^3 I2 f" J* S. W0 F3 b5 m5 @• Unit 7.35 Assessment of Histone Acetylation Levels in Relation to Cell Cycle Phase( H; m( `! r' ]
• Unit 7.36 Click‐iT Proliferation Assay with Improved DNA Histograms6 I- ~! ]* j1 C$ q
• Unit 7.37 High‐Resolution Multiparameter DNA Flow Cytometry for the Detection and Sorting of
; J6 _2 m8 [; y2 Q7 @Tumor and Stromal Subpopulations from Paraffin‐Embedded Tissues
+ C f& ^3 x7 j• Unit 7.38 Dual‐Pulse Labeling Using 5‐Ethynyl‐2′‐Deoxyuridine (EdU) and 5‐Bromo‐2′‐
: ~( w( b) A2 v6 D& {Deoxyuridine (BrdU) in Flow Cytometry- m4 N# H( x5 _* I
• Unit 7.39 High‐Resolution Cell Cycle and DNA Ploidy Analysis in Tissue Samples
; c$ ?+ @6 l2 v( ^1 t+ G• Unit 7.40 Zinc Fixation for Flow Cytometry Analysis of Intracellular and Surface Epitopes, DNA+ R1 @: R5 g- m [
Content, and Cell Proliferation& P! L/ \/ F% P2 v- t
• Unit 7.41 High‐Resolution Cytometry for High‐Content Cell Cycle Analysis/ E: u7 D2 ~ c
• Unit 7.42 Confocal Microscopy for High‐Resolution and High‐Content Analysis of the Cell Cycle8 a" l9 ? X0 c9 N
• Unit 7.43 Application of Click Chemistry Conditions for 5‐Bromo‐2′‐Deoxyuridine Determination
( K5 h& E: p. V: _' J# X. L! _Through Fenton and Related Reactions3 Z: n' o6 c1 n6 v$ `( X/ h: c
• Unit 7.44 Flow Cytometry of Murine Spermatocytes# n) w, n, o: b1 |4 v$ X) G/ [" w
• Unit 7.45 Simultaneous, Single‐Cell Measurement of Messenger RNA, Cell Surface Proteins, and
& y1 j q% c5 A4 jIntracellular Proteins
9 o; X. }: k. O# b• Unit 7.46 Measurement of Low‐Abundance Intracellular mRNA Using Amplified FISH Staining4 a% d* B1 N" W
and Image‐Based Flow Cytometry% a7 Y1 ^7 ]2 a8 u5 Z1 C3 a+ U1 |
• Unit 7.47 Measurement of T‐Cell Telomere Length Using Amplified‐Signal FISH Staining and* n8 F3 ]2 y; k/ z# F$ S
Flow Cytometry
+ w0 y( r: V8 S: `% o• Chapter 8 Molecular Cytogenetics
0 m. {/ p9 W9 [" D9 D4 I• Introduction0 }3 X/ a9 B* Z* N2 ]* o4 ?' F$ G% P: A
• Unit 8.1 Overview of Fluorescence In Situ Hybridization Techniques for Molecular Cytogenetics
/ d3 V p' l* ?• Unit 8.2 Basic Preparative Techniques for Fluorescence In Situ Hybridization
$ ]1 |8 m5 F k# |• Unit 8.3 Probe Labeling and Fluorescence In Situ Hybridization
! `1 G; U$ F4 O# I% q( D• Unit 8.4 Immunocytochemical Detection
! v. X3 K" R& o! h2 u; g5 P• Unit 8.5 Processing and Staining of Cell and Tissue Material for Interphase Cytogenetics
- I+ L" ^9 m. |" E% z- u3 o( t• Unit 8.6 Advanced Preparative Techniques to Establish Probes for Molecular Cytogenetics
8 w. A0 Z4 q- c* A: t# p• Unit 8.7 Combination DNA/RNA Fish and Immunophenotyping
% N+ T, r R5 x/ ~• Unit 8.8 Single‐Nucleotide Sequence Discrimination In Situ Using Padlock Probes6 j1 R+ b" l1 l9 v
• Unit 8.9 Tyramide Signal Amplification (TSA) Systems for the Enhancement of ISH Signals in( i' O5 _5 _1 t
Cytogenetics
S; z t- w0 i4 y& G2 J% P4 k• Unit 8.10 Molecular Combing6 W+ l! p4 R- e6 D. T
• Unit 8.11 Principles and Applications of PRINS in Cytogenetics
; A0 W# `0 v3 w- Q# ?9 t• Unit 8.12 Comparative Genomic Hybridization (CGH)—Detection of Unbalanced Genetic
. s: c3 F2 ]: a! Y7 `% `3 R- E4 vAberrations Using Conventional and Micro‐Array Techniques
- v7 t6 c3 r9 H; `• Unit 8.13 Combined Immunofluorescence and FISH: New Prospects for Tumor Cell5 L1 s' P+ Z7 w7 u, q4 @& [# W
Detection/Identification" f$ J; m. Z- \1 d7 [% D2 n2 F: z
• Unit 8.14 Application of Flow‐FISH for Dynamic Measurement of Telomere Length in Cell
: q* e$ g" B v' pDivision
# f* D* O4 W( n3 `+ n• Chapter 9 Studies of Cell Function
+ f8 b* v7 l- V• Introduction7 T% ?4 d* e' M
• Unit 9.1 Overview of Functional Cell Assays
+ p* J: |8 Q; y3 {3 B) N, u2 B• Unit 9.2 Assessment of Cell Viability& J* k' A% H: O& g- }
• Unit 9.3 Flow Cytometric Measurement of Intracellular pH
$ Z5 t5 l7 a3 O( C+ X; q5 B* _- k4 e• Unit 9.4 Analysis of Intracellular Organelles by Flow Cytometry or Microscopy. f- Y! w- z! x" g
• Unit 9.5 Reporters of Gene Expression: Enzymatic Assays9 H8 L; S7 K. w- v
• Unit 9.6 Estimation of Membrane Potential by Flow Cytometry
; b! p; s g; e# x/ Y- N, _• Unit 9.7 Oxidative Metabolism
$ G* m' P1 S) ?1 D; w• Unit 9.8 Measurement of Intracellular Ions by Flow Cytometry
) z& }; F; p/ | W E9 T6 r• Unit 9.9 Intracellular Cytokines
1 m" r; G6 o# @' r' K• Unit 9.10 Assays of Natural Killer (NK) Cell Ligation to Target Cells
2 T: [- k" Z. o# |% l7 z• Unit 9.11 Flow Cytometric Analysis of Cell Division by Dilution of CFSE and Related Dyes% C5 d% C3 R. x5 O
• Unit 9.12 Fluorescent Proteins for Flow Cytometry# g5 H" s$ m. I( ^- m
• Unit 9.13 In Vitro Invasion Assays: Phagocytosis of the Extracellular Matrix
$ T5 d4 p: \2 q' J• Unit 9.14 Flow Cytometric Analysis of Mitochondrial Membrane Potential Using JC‐18 j. W, r3 l- U2 \
• Unit 9.15 Multiparameter Analysis of Physiological Changes in Apoptosis
0 C" [; r+ I; H! G- @5 T( _* H. s• Unit 9.16 Signal Transduction During Natural Killer Cell Activation
+ k9 D# s( I. W9 f1 D; j• Unit 9.17 Assessment of Surface Markers and Functionality of Dendritic Cells (DCs)4 [4 ~# g0 w+ f" i
• Unit 9.18 Stem Cell Identification and Sorting Using the Hoechst 33342 Side Population (SP)/ Z" o% X+ l: u
• Unit 9.19 Assessment of Phagocyte Functions by Flow Cytometry
* \8 `4 {! h# k4 O* x, P, E• Unit 9.20 Flow Cytometric Analysis of Calcium Mobilization in Whole‐Blood Platelets
) j0 E' g/ |6 L3 V: M6 l# }7 I• Unit 9.21 Flow Cytometric Analysis of Cytokine Responses in Stimulated Whole Blood:
& c* A( x- `; H& [9 l- |& O+ fSimultaneous Quantitation of TNF‐α‐Secreting Cells and Soluble Cytokines
5 ~8 v- p% X, D% J% S! T/ _) U• Unit 9.22 Optimized Whole‐Blood Assay for Measurement of ZAP‐70 Protein Expression
* _* [7 u' Z. J' m1 @• Unit 9.23 Flow Cytometry of the Side Population (SP)
& G9 @% M" P" ]• Unit 9.24 High‐Throughput Cytotoxicity Screening by Propidium Iodide Staining
0 H) F6 H0 C6 ?; e3 L• Unit 9.25 Advanced Application of CFSE for Cellular Tracking
9 ]( w6 A9 }. e$ ]4 b• Unit 9.26 Immunophenotyping and DNA Content Analysis of Acetone‐Fixed Cells
8 ~: u5 T3 ], ?! Y7 C• Unit 9.27 Whole Blood Processing for Measurement of Signaling Proteins by Flow Cytometry/ _+ C7 F, T; Q. p0 j1 `3 T2 [
• Unit 9.28 Measurement of Cytoplasmic to Nuclear Translocation# {& c% x* B" K* J" M/ N
• Unit 9.29 Overview of Very Small Embryonic‐Like Stem Cells (VSELs) and Methodology of Their5 x% h f, R$ _" n" E7 _
Identification and Isolation by Flow Cytometric Methods, A [ \ i7 Q: W1 G
• Unit 9.30 Stem Cell Side Population Analysis and Sorting Using DyeCycle Violet2 Q+ F8 o# R* c- T
• Unit 9.31 Measurement of Phagocytosis and of the Phagosomal Environment in# r! G4 n2 _8 g: ]% } V
Polymorphonuclear Phagocytes by Flow Cytometry
3 y, [6 e7 \! o0 s9 U" h% G• Unit 9.32 Yeast Cell Cycle Analysis: Combining DNA Staining with Cell and Nuclear Morphology
' k8 P- ~- `4 t, r• Unit 9.33 Identification of Endothelial Cells and Progenitor Cell Subsets in Human Peripheral& H( F% d0 ^' X& M5 F! N: @
Blood8 s5 r" B9 ~- }+ f7 O
• Unit 9.34 Amine‐Reactive Dyes for Dead Cell Discrimination in Fixed Samples
( z* E8 y2 E$ _1 B" o• Unit 9.35 Detection of Intracellular Glutathione Using ThiolTracker Violet Stain and
, t4 A- ]& r/ y; a/ i! J0 w. T3 fFluorescence Microscopy
% K. p9 t7 l' {9 \: m• Unit 9.36 In Situ Proximity Ligation Assay for Microscopy and Flow Cytometry. P3 b3 p- l1 R% p; ?( A
• Unit 9.37 Assessing Mitochondrial Redox Status by Flow Cytometric Methods: Vascular" r% t& H0 j9 R# W9 V7 s! b' h
Response to Fluid Shear Stress
" L Z! F' P' e6 u- g• Unit 9.38 A Violet Ratiometric Membrane Probe for the Detection of Apoptosis
3 T' `# V6 n* ^• Unit 9.39 Ex Vivo Imaging of Excised Tissue Using Vital Dyes and Confocal Microscopy; ?3 f; X+ x8 }; ]- S+ H$ I
• Unit 9.40 Flow Cytometry‐Based Quantification of Cell Proliferation in the Mixed Cell Co‐Culture @9 Q+ ?$ U3 b4 G; k
• Unit 9.41 Kinetic Viability Assays Using DRAQ7 Probe8 E0 h, \6 d* N. E: p
• Unit 9.42 Multiparameter Analysis of Apoptosis Using Lab‐on‐a‐Chip Flow Cytometry4 K, M1 L. G# w9 S. N
• Unit 9.43 Real‐Time Detection of Protein Trafficking with High‐Throughput Flow Cytometry
! ~* H9 Q1 j8 g! G8 o- _: r4 w(HTFC) and Fluorogen‐Activating Protein (FAP) Base Biosensor+ B, M2 r! q& U- x
• Unit 9.44 OpenSource Lab‐on‐a‐Chip Physiometer for Accelerated Zebrafish Embryo Biotests
4 `/ d5 T& a! Y! H7 m1 |" B1 D6 T• Unit 9.45 Measurement of Autophagy by Flow Cytometry& i1 E& v! R( V
• Unit 9.46 Immunophenotyping of Paucicellular Samples
! {4 _% w* Q( i• Unit 9.47 Attenuation of Replication Stress–Induced Premature Cellular Senescence to Assess- j$ t Z/ v# S# _# Y2 f% v( j
Anti‐Aging Modalities
1 b* l7 x; Q* o3 P• Unit 9.48 High Throughput‐Based Mitochondrial Function Assays by Multi‐Parametric Flow6 ]. N- _7 Y, C* ~8 P
Cytometry: n* N! o+ l7 I8 n+ E1 X' E- R
• Unit 9.49 Measurement and Characterization of Apoptosis by Flow Cytometry& L# i1 L+ e/ Z; l
• Unit 9.50 Identification of Human Memory‐Like NK Cells
' s/ ^# V- V+ f. l" B8 j' J( c9 Q• Unit 9.51 Quantitative Analysis of Cellular Senescence in Culture and In Vivo
# k+ K) l# c( I6 d& F7 J j• Unit 9.52 Method to Detect the Cellular Source of Over‐Activated NADPH Oxidases Using, X* w5 B3 s( Z
NAD(P)H Fluorescence Lifetime Imaging
( B' h/ Y, b% p3 C3 ?5 E, M• Chapter 10 Data Processing and Analysis
3 _1 R0 o5 r" k) |& |0 f% y• Introduction6 r* ?4 v& ^+ u9 X
• Unit 10.1 Data Management
$ }1 k8 P% m" q• Unit 10.2 Data File Standard for Flow Cytometry, FCS 3.0
/ c' S% h3 Z- a; `% E! p$ M0 K• Unit 10.3 Listmode Data Processing
( k. o4 p/ i, W- `• Unit 10.4 Multidimensional Data Analysis in Immunophenotyping
% n, u( o; C% b0 s/ B J• Unit 10.5 Two‐Dimensional Image Processing and Analysis
* z% ]) P7 `. G" d* V5 ^• Unit 10.6 Data Presentation3 R+ d6 B/ ^) L0 r: p
• Unit 10.7 Data Analysis Through Modeling
0 ~5 [& `9 \- t F• Unit 10.8 Multivariate Analysis
% A& D; j$ a! B2 _6 r7 B8 o' P- Q• Unit 10.9 Detection and Location of Hybridization Domains on Chromosomes by Image
1 K* J7 i! l8 cCytometry
9 b6 Q% {% o3 D• Unit 10.10 Three‐Dimensional Image Visualization and Analysis# l/ b( z& X6 O$ ]
• Unit 10.11 Image Processing and 2‐D Morphometry
2 V$ }4 P. D* B+ m b4 Z• Unit 10.12 Dial‐In Flow Cytometry Data Analysis( ~3 e! ~. E5 I
• Unit 10.13 The Application of Data Mining to Flow Cytometry% x4 d& z, B# r
• Unit 10.14 Intensity Calibration and Flat‐Field Correction for Fluorescence Microscopes
1 E$ _6 Q u4 i# g3 {* ~2 u2 ]4 x• Unit 10.15 A Software Method for Color Compensation) d* q5 Z+ J5 v' ]
• Unit 10.16 Alternatives to Log‐Scale Data Display
6 m: r) i) g& e5 Q" k# q! c• Unit 10.17 Web‐Based Analysis and Publication of Flow Cytometry Experiments
. E. _7 s- F4 x3 V2 H% m% e7 v• Unit 10.18 Preparing a Minimum Information about a Flow Cytometry Experiment (MIFlowCyt)6 ?" D+ q+ U6 D8 k
Compliant Manuscript Using the International Society for Advancement of Cytometry (ISAC)2 r8 c6 R( o& t1 `, p+ A! `
FCS File Repository (FlowRepository.org)3 C% w& g* Q5 E- k, E
• Unit 10.19 Digital Data Acquisition and Processing# I. I6 j1 f; D) K: }( ]' t% S4 C7 p
• Chapter 11 Microbiological Applications
% S4 S1 q/ Y0 x& g) e7 b4 Z• Introduction
& \9 [" V9 @5 l+ s' |$ X! E; A• Unit 11.1 Overview of Flow Cytometry and Microbiology
4 j. v7 k* W1 l+ F& x• Unit 11.2 Flow Cytometry and Environmental Microbiology0 F0 i+ b$ P7 z0 `2 U
• Unit 11.3 Estimation of Microbial Viability Using Flow Cytometry
8 E8 [+ _$ k: E f" J! e4 C2 d• Unit 11.4 Sorting of Bacteria2 }. w7 p8 F- Z1 K, d* q1 R9 Y' u' L
• Unit 11.5 Detection of Borreliacidal Antibodies by Flow Cytometry
]( f( `9 S$ {7 W- z D* G: s• Unit 11.6 Flow Cytometric Detection of Pathogenic E. coli in Food2 m2 O3 N) }3 w- z2 Q
• Unit 11.7 Mycobacterium tuberculosis Susceptibility Testing by Flow Cytometry8 | l! K9 W* ^! O/ |
• Unit 11.8 Antibiotic Susceptibility Testing by Flow Cytometry
% z8 F7 a, w3 O0 W; U• Unit 11.9 Determination of Bacterial Biomass from Flow Cytometric Measurements of Forward
* D+ P% [9 f* V) n' W1 d1 o- `/ SLight Scatter Intensity
5 s' ~. _& C1 V. q• Unit 11.10 Flow Cytometry of Yeasts
8 j9 W# { F& ^9 G( S• Unit 11.11 Enumeration of Phytoplankton, Bacteria, and Viruses in Marine Samples
6 c1 k8 h9 `$ B6 y4 \• Unit 11.12 DNA/RNA Analysis of Phytoplankton by Flow Cytometry5 `, y' \7 [5 h, L% A$ u
• Unit 11.13 Cell Cycle Analysis of Yeasts
2 X! ?" [3 z% k, B1 L- k• Unit 11.14 Flow Cytometric Assessment of Drug Susceptibility in Leishmania infantum& E" c7 m' ]" [# e+ S4 L
Promastigotes4 s, A8 O5 W8 i
• Unit 11.15 Resolution of Viable and Membrane‐Compromised Free Bacteria in Aquatic
0 i6 K+ R) Y4 x; gEnvironments by Flow Cytometry* {, Y7 P& ?+ k- Z0 E: v# ?
• Unit 11.16 Functional Assays of Oxidative Stress Using Genetically Engineered Escherichia coli0 x3 C1 m- C9 R) a* v3 n D- S
Strains
% s( @8 C i0 X( \8 _3 r. b• Unit 11.17 Labeling of Bacterial Pathogens for Flow Cytometric Detection and Enumeration
5 [1 u5 g4 H' {• Unit 11.18 Detection of Extracellular Phosphatase Activity of Heterotrophic Prokaryotes at the5 ^9 F" M) K6 z1 X/ P: U' }
Single‐Cell Level by Flow Cytometry- J8 k9 { o: i6 s N. ^' e2 X
• Unit 11.19 Life Cycle Analysis of Unicellular Algae
' D5 h- }, [9 g, y @• Unit 11.20 Cytometry in Malaria—A Practical Replacement for Microscopy?9 j9 f4 O2 w5 }2 J
• Unit 11.21 Large Particle Sorting to Isolate Live Parasitic Nematode Eggs# H) Y8 n7 D; Z3 l
• Unit 11.22 High‐Throughput Particle Uptake Analysis by Imaging Flow Cytometry! X2 M0 S5 o S8 z+ q% h& {
• Chapter 12 Cellular and Molecular Imaging( D" _4 D0 F. J/ Q
• Introduction" |4 Z1 l; r) O$ W, Z& B5 m
• Unit 12.1 Comparative Overview of Flow and Image Cytometry" N# O9 l9 L; {- z
• Unit 12.2 Basics of Digital Microscopy
/ m, A! }$ f: }: t• Unit 12.3 Modern Confocal Microscopy
/ h# ]5 B2 S, D3 ?( u9 h! y. B, o• Unit 12.4 Time‐Lapse Microscopy Approaches to Track Cell Cycle and Lineage Progression at
/ t/ L# c5 n- d& C! L) j. P/ o. Gthe Single‐Cell Level
6 Y5 {# U( C7 {/ t$ Y; p; `• Unit 12.5 Three‐Dimensional Visualization of Blood and Lymphatic Vasculature in Tissue Whole( q8 m! b3 ?+ r P# ?
Mounts Using Confocal Microscopy& L; g3 x: v4 |6 l% i. f1 Z
• Unit 12.6 Quantitative Fluorescence In Situ Hybridization (QFISH) of Telomere Lengths in! ^% j+ J$ }0 s& q
Tissue and Cells( I( u* R& |& F" L
• Unit 12.7 Detecting Protein–Protein Interactions with CFP‐YFP FRET by Acceptor Photobleaching; X4 p* G# ~/ Y6 ]& J" q3 y
• Unit 12.8 Measuring FRET in Flow Cytometry and Microscopy
4 J- b; Y- b$ c, J9 E) f• Unit 12.9 Live‐Animal Imaging of Renal Function by Multiphoton Microscopy [, l; R% ^9 L
• Unit 12.10 Detecting Protein‐Protein Interactions In Vivo with FRET using Multiphoton
& i. S" O) h8 |# g8 zFluorescence Lifetime Imaging Microscopy (FLIM)
, f0 w! x/ ~# W* @+ A; ^4 J( Z u1 i3 S• Unit 12.11 Confocal Imaging of Cell Division
& Z, l2 W3 S% p0 Y8 i• Unit 12.12 From In Vitro to In Vivo: Imaging from the Single Cell to the Whole Organism
, O( S8 u$ w6 y2 t% {3 Z• Unit 12.13 Use of Spectral Fluorescence Resonance Energy Transfer to Detect Nitric Oxide‐
0 V/ Q t- R# n P( @2 ~' d9 IBased Signaling Events in Isolated Perfused Lung
) n: N& c4 W; X% M6 k( E9 n• Unit 12.14 Flow Cytometric FRET Analysis of ErbB Receptor Tyrosine Kinase Interaction8 m% z s; f' l9 _
• Unit 12.15 Cryosectioning
0 {6 u. C, Y9 q8 y8 u; j% |- [% @6 _• Unit 12.16 Immunohistochemistry7 y8 m, C1 K2 y+ U4 Q! M( K2 Q9 n
• Unit 12.17 Simultaneous Optical Mapping of Intracellular Free Calcium and Action Potentials
2 f' ? ^ ^ ^from Langendorff Perfused Hearts' p3 U' D* D. C2 j! `7 X
• Unit 12.18 Total Internal Reflection Fluorescence (TIRF) Microscopy
5 F, Y& z, ^9 a4 s( d# [• Unit 12.19 3D Deconvolution Microscopy
% X3 X5 Z6 _% w u9 h9 |7 w• Unit 12.20 Approaches to Spectral Imaging Hardware
9 c, Q3 E, M' m' Y• Unit 12.21 From Image to Data Using Common Image‐Processing Techniques# F( A! e. u" }! X4 H, r* ^- U
• Unit 12.22 Setting Up and Running an Advanced Light Microscopy and Imaging Facility [; |/ E, ^( Z5 d% W7 a
• Unit 12.23 Photoactivation and Imaging of Optical Highlighter Fluorescent Proteins P5 ^' f8 G2 `) C
• Unit 12.24 Practical Methods for Molecular In Vivo Optical Imaging
" E7 y- f! L, y• Unit 12.25 Characterization of Surface FAS—Quantitative Morphological Analysis Using
; k5 U/ e; C7 Q( C; S9 X* VQuantitative Imaging Cytometry C( N, P0 Q! F& X
• Unit 12.26 Two‐Photon Imaging of the Immune System5 X; Z. P: D g
• Unit 12.27 Near‐Infrared Molecular Probes for In Vivo Imaging* Q2 s0 T2 `- V0 b# q) D# v
• Unit 12.28 Live Imaging of the Lung
2 p: i: f7 [, Q; y9 x) v5 \, L• Unit 12.29 Total Internal Reflection Fluorescence (TIRF) Microscopy Illuminator for Improved) b$ t& D$ E% Q) A
Imaging of Cell Surface Events
& Q' R# U5 a" c6 K* s9 u: J. R• Unit 12.30 A Review of Reagents for Fluorescence Microscopy of Cellular Compartments and
( B8 k, @" J7 iStructures, Part I: BacMam Labeling and Reagents for Vesicular Structures9 J6 Y+ B% i& f( O- C4 P$ v
• Unit 12.31 A Review of Reagents for Fluorescence Microscopy of Cellular Compartments and3 O; L, S6 f _6 n5 p6 S7 I4 [2 b# Z; F
Structures, Part II: Reagents for Non‐Vesicular Organelles
& q6 V/ Q' D( O• Unit 12.32 A Review of Reagents for Fluorescence Microscopy of Cellular Compartments and
" Q8 u+ t" B2 ~6 [4 OStructures, Part III: Reagents for Actin, Tubulin, Cellular Membranes, and Whole Cell and
; |3 d" h' k9 JCytoplasm: T; x4 j- }; F) L7 ~( a6 j
• Unit 12.33 A Rapid and Sensitive Automated Image‐Based Approach for In Vitro and In Vivo( Z, }/ @- N/ D. `
Characterization of Cell Morphology and Quantification of Cell Number and Neurite Architecture: @, P% l: [$ }) G. m0 \3 F
• Unit 12.34 Imaging Autophagy
, u/ z7 N: y; G$ K: |) i& H( z• Unit 12.35 The Application of KillerRed for Acute Protein Inactivation in Living Cells+ a! j2 a5 A+ c# S: W
• Unit 12.36 Correlative Fluorescence and Electron Microscopy
, O- q( ^, x/ A" K) x% C• Unit 12.37 Light Sheet Fluorescence Microscopy (LSFM)- T$ B M* p( A* k3 U+ _
• Unit 12.38 Semi‐Automated Object Tracking Methods in Biological Imaging
6 a9 o( |) [$ E( g6 x• Unit 12.39 Cell Volume Measurements by Optical Transmission Microscopy
^/ K S/ |& B• Unit 12.40 Visualization of Telomere Integrity and Function In Vitro and In Vivo Using
. b% g3 R& n# c. T/ y, Y' C' fImmunofluorescence Techniques/ B W' I# C0 V s) V
• Unit 12.41 Microscopic Investigation of Protein Function in C. elegans Using Fluorescent8 S* j( P2 Q1 L
Imaging
3 n# _$ e: G+ n1 R• Unit 12.42 In Vivo Immuno‐Spin Trapping: Imaging the Footprints of Oxidative Stress
3 z9 q1 Y4 t& n& A( r• Unit 12.43 High‐Content Microscopy Analysis of Subcellular Structures: Assay Development
- W) ]6 I1 e# nand Application to Focal Adhesion Quantification2 {3 i3 {% p# O8 _6 D2 U; f$ P: C3 q
• Unit 12.44 Automated Measurement of Blood Vessels in Tissues from Microscopy Images& |( k% N" w" O& j: T: G1 s% x
• Unit 12.45 Correlative Fluorescence and Electron Microscopy in 3D—Scanning Electron( }! U9 C, t3 n6 T* K
Microscope Perspective
, {6 i# m$ F$ q2 |3 v+ M3 z% x• Chapter 13 Multiplexed and Microparticle‐Based Analyses6 H7 ]$ X3 q% U/ P' J. |; ^3 W
• Introduction3 H+ w" Z' G4 A) V) v; S
• Unit 13.1 Multiplexed Microsphere‐Based Flow Cytometric Immunoassays
1 T. J3 ?% o( v9 J1 p5 d& G1 {• Unit 13.2 Microsphere Surface Protein Determination Using Flow Cytometry
- s/ E; f5 a/ s' A' D• Unit 13.3 Use of Microsphere‐Supported Phospholipid Membranes for Analysis of Protein‐Lipid
8 S9 i: c' S2 w- d$ wInteractions
% o! l* u, a' `. h: U3 l, T• Unit 13.4 Multiplexed SNP Genotyping Using Primer Single‐Base Extension (SBE) and
6 g0 X3 g* i) k# zMicrosphere Arrays. V; y# P. n3 D$ L: [( |5 T
• Unit 13.5 BeadCons: Detection of Nucleic Acid Sequences by Flow Cytometry, X6 A/ n& c2 x
• Unit 13.6 Characterization of Nuclear Receptor Ligands by Multiplexed Peptide Interactions
l5 F: g7 {. d( J7 S* J9 m• Unit 13.7 Detection of Gene Fusions in Acute Leukemia Using Bead Microarrays9 M @3 G) F4 B* E! l
• Unit 13.8 Reagents and Instruments for Multiplexed Analysis Using Microparticles8 u& r4 {( {+ ]) o8 s- G
• Unit 13.9 Multiplexed Detection of Fungal Nucleic Acid Signatures0 E- L6 x+ D$ |( V+ p$ k* w J
• Unit 13.10 Multiplexed Analysis of Peptide Antigen‐Specific Antibodies
0 n2 m' ]# }$ n3 O! E% t2 E+ Y• Unit 13.11 Use of Flow Cytometric Methods to Quantify Protein‐Protein Interactions
. ?8 u. O/ X" ~$ F. A: t• Unit 13.12 Microsphere‐Based Flow Cytometry Protease Assays for Use in Protease Activity8 P+ t) T! k6 b: ^. ?6 Y$ Y
Detection and High‐Throughput Screening
9 u. m: X; @. X- P• Unit 13.13 Application of the PrimRglo Assay Chemistry to Multiplexed Bead Assays W0 y, ], t M3 ]; |. A; E
• Unit 13.14 Flow Cytometry of Extracellular Vesicles: Potential, Pitfalls, and Prospects: J4 w4 |, a6 P6 M1 Z P7 [
• Unit 13.15 Optimized MOL‐PCR for Characterization of Microbial Pathogens, k9 l$ T7 p3 F) O- U
• Appendix 1 Abbreviations and Useful Data- F# ?9 e, F2 T& Q- H' ^
• Appendix 1A Abbreviations Used in this Manual' W9 z4 E, }8 P( {* ^8 P$ r% N
• Appendix 1B Common Conversion Factors
( v5 u( b1 \( D d% H t• Appendix 2 Stock Solutions, Equipment, and Laboratory Guidelines
# K' e- S1 ?( z7 M8 T) R a• Appendix 2A Common Stock Solutions, Buffers, and Media
, W, G V K4 @• Appendix 3 Commonly Used Techniques# V; t' ~1 t. m2 H5 a8 d# a" ~
• Appendix 3A Cell Counting0 s! H) G# T$ X' ]
• Appendix 3B Techniques for Mammalian Cell Tissue Culture) ^7 S `) o0 q9 W
• Appendix 3C Diagnosis and Treatment of Mycoplasma‐Contaminated Cell Cultures
2 f4 ?4 E' u' R• Appendix 3D Wright‐Giemsa and Nonspecific Esterase Staining of Cells
3 {0 K' R- G; [, I7 R3 s• Appendix 3E Techniques for Bacterial Cell Culture: Media Preparation and Bacteriological Tools) B' m f9 X" H6 Q: w3 P
• Appendix 3F Growing Bacteria in Liquid Media
4 K d3 r3 i/ Q" @ T. z• Appendix 3G Growing Bacteria on Solid Media4 b) f6 M# N; N& @: R; O- a: j) Q
• Appendix 3H Importing Biological Materials! m4 R4 h4 W1 ]% O: G5 z
• Appendix 3I Production of Polyclonal Antisera
: u- E9 S7 P- w* Y$ |• Appendix 3J Production of Monoclonal Antibodies" H/ j; e2 x: I# F3 I5 ]0 N! u
• Appendix 3K Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization |
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发表于 2017-6-6 03:16
