Identification of Novel Sperm Surface Markers via HTS - Background

omarys's version from 2017-08-10 17:03

Section 1: Introduction

Question Answer
The human cell surface marker screening panel isa collection of cell surface marker antibodies
The human cell surface marker screening panel is designed toscreen cells for the expression of 373 human cell surface proteins
The human cell surface marker screening panel employs which biotechnology?flow cytometry
The significance of these panels is their potential for developing new strategies tocharacterize and isolate sperm from ejaculate of azoospermic patients (or from testicular biopsy)
There are ___ protein expression profiles at ___ states of differentiation.different ; various
HTShigh-throughput screening (and HTFC is flow cytometry)
The parameters measured in flow cytometry are- (3)fluorescence intensity (up to 18 colors); forward scatter; side scatter

Section 2: What's flow cytometry, precious? pt. 1

Question Answer
FC is a ___-based technology.laser
Applications are- (4)cell counting; cell sorting; detection of biomarkers; protein engineering
FCcharacterizes and defines different cell types in a heterogeneous cell population
FC also analyzes cell ___ and ___.size ; volume
FC is used to measure ___ ___ produced by fluorescent-labeled ___ detecting proteins, or ___ that bind to specific cell-associated molecules.fluorescence intensity ; antibodies ; ligands
In the cytometer, cells/particles passing through the laser beam ___ light, which is detected as ___ and ___.scatter ; forward scatter ; side scatter
FS correlates with ___ ___ and SS is proportional to the ___ of the cellscell size ; granularity (so using FS and SS alone helps differentiate cell size and granularity)
When running a blood sample thru the cytometer, granulocyte cells produce ___ FS and ___ SS; monocytes produce ___ FS and ___ SS; lymphocytes produce ___ FS and ___ SS.high; high ; high; low ; low; low
A graph used in FC is adot plot of FS versus SS

Section 3: Flow cytometry pt. 2

Question Answer
The analysis software used in the project isBD FACSDiva (BD FACSCanto II is for obtaining flow cytometry data)

Section 4: Project details

Question Answer
HTP flow cytometric analysis was carried out on ___ sperm from 4 donors (carried out to screen for the presence of 373 receptors).healthy/fertile
Among the 373 receptors, IL7-R (___) had low prevalence in healthy sperm samples. This receptor is known for its ___ function.CD127 ; anti-apoptotic
In asthenozoospermic samples, there was a significant ___ in IL7R+ve spermatozoa.decrease
Aim #1 was to identify new extracellular markers for the isolation of rare ___ spermatozoa in ___ ___, thus avoiding ___. (Aim #2 was to do the same in NOA ___ ___.)functional ; NOA ejaculate ; mTESE (testicular biopsy)
CD52 (expressed on lymphocytes and epididymal epithelial cells) is AKAmaturation-associated sperm antigen
In one study, it was found that CD52 is ___ ___ for fertilization in the mouse, either _________ or _________.not required ; in-vivo ; in-vitro

Section 5: Andrology update: TEX101

Question Answer
An imaging flow cytometerAmnis ImageStream (optimal for 8 colors...max is 11)
Objective #1 of project is to confirm that testicular sperm have 2 ___ ___ and identify the ___ of the sperm contained within the ___.haploid peaks ; morphology ; peaks
Objective #2 is tot