细胞功能测定

Protocol I: Triton X-100 Lysis Buffer In 96 flat-wells plate incubate 4x10 6 target cells (40 wells of 105 per well) with desired concentration of effectors (105 target cells per well). After incubati ...

1. Introduction Nuclear morphology changes characteristic of apoptosis appear within the cell together with a distinctive biochemical event: the endonuclease-mediated cleavage of nuclear DNA. In fact ...

I. Protocol1. Harvest cellsOptional: wash plate with 37°C PBS (gently so as not to lose cells); check on microscopeafter aspirate PBS or mediaPlace plate on iceLyse with DNA ladder buffer 4°C150 ul/60 ...

Protocol for Frozen Sections:Warm 150ml 4% Paraformaldehyde/1x PBS to RT. Fix slides in it 20 min. RT. 1x PBS rinse 2 times. 1x PBS 30 min. RT. Begin chilling Triton/SSC on ice. 0.1% Triton/ 0.1% Sodi ...

1. INTRODUCTION Apoptosis was observed from invertebrates to lower and higher verterbrates and intervenes both in physiological and in pathological states such as normal cell turnover embryogenesis ...

Materials8% (w/v) paraformaldehyde stock solution: Dissolve 8 g of powdered paraformaldehyde in 100 ml water. Heat and stir (55-60 C � do not go higher). Add a few drops of 2 N sodium hydroxide until ...

When an irreplaceable culture becomes contaminated researchers may attempt to eliminate or control the contamination. First determine if the contamination is bacteria fungus mycoplasma or yeast. Isola ...

Purpose: This method is advantageous for saving the occasional cultures that become contaminated. Yeast contaminated cultures will appear cloudy when slightly shaken and lymphocytes will not cluster t ...

DescriptionThis protocol is used in our lab to reduce the costs of the cell sorting with MACS reagents. The cell suspension obtained after this protocol contains 40-70% monocytes. This cell suspension ...

This protocol describes a procedure for isolating human peripheral blood mononuclear cells (lymphocytes and monocytes) from a Buffy Coat (obtained from a blood bank). Platelets red blood cells and neu ...

1. Dilute 2 ml of 4000 U/ml Collagenase D as follows: 1 ml into 9 ml HBSS/Ca++/Mg++ (=400U/ml) and 1 ml into 39 ml HBSS/Ca++/Mg++ (=100U/ml). Put on ice. 2. Place 5 ml of the 100U/ml solution in a 10 ...

Freezing and Thawing cells Freezing It is best to freeze cells that are growing rapidly. With adherent cells it is easiest to set up 100 mm dishes give them fresh medium the day before you freeze them ...

1) Keep prepared solutions on ice.2) Determine total cell count of cells to be frozen. (e.g. 1 X 108 )3) Determine number of vials to be frozen. (e.g. 10 vials at 1 X 107 )4) Centrifuge cell suspensio ...

Accuracy of manual counts with an hemacytometer depend on: accurate mixing of the sample (no bubbles!) number of chambers counted number of cells counted (practical: 200-500 per 0.1 mm3) The chamber o ...

1) Turn on the counter by pulling out the on/off button. You need to do this at least 10 min before use to obtain sufficient vacuum.Usually put 0.2 ml of your suspension of cells into 10 ml Tris or TD ...

OUTLINE We consider manual counts as less quick but more precize than authomatic (electronic) since in the latter case the counts are based on the size of particles so everything that has the same siz ...

Method: Cell Counts Using a Hemacytometer June 1 1990 Rosalie Veile Purpose: The purpose of this procedure is to determine the cell density of the culture. Cell cultures always have some dead cells; t ...

A hemacytometer (also spelled hemocytometer) is an etched glass chamber with raised sides that will hold a quartz coverslip exactly 0.1 mm above the chamber floor. The counting chamber is etched in a ...

There are many procedures with which to trypsinize cells. All include washing the cell monolayer with TD or in rare cases with VE. This removes serum which contains trypsin inhibitors and lowers the c ...

Method: Lymphoblastoid Cell Lines from Frozen Whole BloodMay 31 1990 Rosalie Veile Purpose: Blood Samples can be stored frozen as a backup in case an LCL is needed at a later date.Time required: 15 mi ...