Mousekeratinocytecultures
PRIMARY MOUSE KERATINOCYTE CULTURESIsolation of epidermal keratinocytes from neonatal mice is based on the protocol of Dlugosz et al., Methods Enzymol. 254:3-20 (1995). The epidermis from a newborn mouse should yield approximately 5-10x106 cells, with a 30-40% plating efficiency.Sacrifice newborn mice by CO2 narcosis. Note: Newborn mice may require 10-15 min in the CO2 chamber.Sterilizin......阅读全文
Mouse－keratinocyte－cultures
PRIMARY MOUSE KERATINOCYTE CULTURESIsolation of epidermal keratinocytes from neonatal mice is based on the protocol of Dlugosz et al., Methods Enzymol
Keratinocyte－Differentiation
The epidermis, which provides a protective barrier that undergoes a constant renewal, is a multi-layered tissue with the proliferating cells located i
Effects－of－calcineurin－in－Keratinocyte－Differentiation
The differentiation of keratinocytes constantly replenishes the upper layers of human skin we lose each day. One factor that contributes to terminal k
Cryopreservation－of－cell－cultures
1. Examine all flasks by inverted-phasemicroscopy. Cultures used for preservation should be grown free of antibiotics, show no signs of microbial cont
ORGANOTYPIC－LIMB－CULTURES
-embryos are dissected from timed-pregnant mice from 10.5 - 11.5 d.p.c.-limb buds are microdissected and placed in holding medium (L15 medium suppleme
Preserving－yeast－cultures
Short term storageYeast cultures are stable for 1-2 weeks when refrigerated. Petri dishes should be sealed or in plastic bags.Medium term storageYeast
Growing－Overnight－Cultures
1. Place 2 mL of the appropriate sterile medium in a 13 mm yellow-capped culture tube. If more culture is needed, place up to 5 mL in a 16 mm green-ca
Hippocampal－Neuron－Cultures
实验概要The protocol provides a method of hippocampal neuron cultures.主要试剂Begin by timing the pregnant mouse at E17-E19 days of gestation. Have ready the
ORGANOTYPIC－KIDNEY－CULTURES
-embryos are dissected from timed-pregnant mice from 11.5 d.p.c. to 13.5 d.p.c.-metanephroi and associated ureteric buds are microdissected and placed
Mouse－Spleenectomy
OUTLINEPROTOCOLGeneral anesthesia1. inject i.p. 250µl/mouse of the ANESTHETIC mixture < wait for 2-10min for the anesthetic effect>2. for verify the a
Transfer－of－Eukaryote－Suspension－Cultures
MaterialsFibroblast suspension cultureTissue culture laminar flow hoodMedia appropriate to culture line usedDisposable pipettes (10 ml and 1.0 ml)Disp
AMNIOTIC－FLUID－CULTURES－ON－COVERSLIPS
I. Purpose:Amniotic fluid may be used for prenatal diagnosis of aneuploidy or other structural abnormalities. II. Culture Procedure:A. Aseptic techniq
Primary－Cultures－fo...
实验概要The following protocol provides a method of primary cultures for IHC – viability assays.实验步骤1. Preparation of primary mesencephalic cultures 1)
Dissociated－Cultures－of－Cerebellar－Neurons
Dissociated Cultures of Cerebellar NeuronsHank Dudek (617-355-4735)Protocolisolate cerebella (ÒCbÓ)cut off head into plate with HHGNhold nose with lar
AMNIOTIC－FLUID－CULTURES－ON－COVERSLIPS
实验概要 AMNIOTIC FLUID CULTURES ON COVERSLIPS主要试剂Solutions:Colcemid working solution: 10 mcg/ml Colcemid in Hank's Balanced Salt Solution, sto
Pathology－and－Autopsy－of－a－mouse
if mouse is found deadif mouse is a newborn or < 1 wk of ageIf mouse is alive or moribund and above one week of ageRoutine dissectionFormalin fixation
Preparation－of－Mouse－Neutrophils
实验概要Preparation of Mouse Neutrophils实验步骤Mice：8-16 weeks old malePrewarm buffer to room temperatureBuffer A : Ca2 /Mg2 ‐free Hank’s buffered saline so
Isolation－of－mouse－embryos
1. Sacrifice impregnated mouse.2. Dissect out the uterus of the mouse. Pulling up on the uterus with one set of forceps,use another to tear the mesome
Preparation－of－Mouse－Neutrophils
实验步骤Mice：8-16 weeks old malePrewarm buffer to room temperatureBuffer A : Ca2 /Mg2 ‐free Hank’s buffered saline solution [HBSS; Invitrogen, Grand Isla
Complete－Mouse－Necropsy
EuthanasiaEuthanasia and mouse necropsies require prior IACUC approval. The mode of euthanasia should be chosen which minimizes pain or distress to th
SOLID－TUMOR－CULTURES－FOR－CHROMOSOME－ANALYSIS
I. Purpose:A. Samples of solid tumors or lymph nodes may be sent from patients with cancer. These samples should be processed directly and also set up
Rodent－Retinal－Ganglion－Cell－Cultures
实验概要Central neurons lose the ability for axonal regrowth during development and typically do not regenerate their axons following axotomy once the
TISSUE－FIBROBLAST－CULTURES－FOR－CHROMOSOME－ANALYSIS
I. Purpose:A: Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconclusive, e.g. suspected
TISSUE－FIBROBLAST－CULTURES－FOR－CHROMOSOME－ANALYSIS
实验概要 Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconclusive, e.g. suspect
Placental－trophoblast－and－chorionic－cell－cultures
1. Placental trophoblast and chorionic trophoblast cells were prepared using a modification of the method. 2. Term human placentae and chorion tissue
Culturing－Mouse－Embryonic－Fibroblasts
MaterialsTrypsin (Gibco 25200-023)3T3 Medium: 500 mL DME (Invitrogen) + 50 mL FBS (Hyclone) + 5 mL 100x Pen/Strep2x Freezing Medium: 3T3 Medium + 20%
Chick－or－Mouse－embr...
实验概要The following procedure describes the procedure for whole mount staining of chick or mouse embryo’s. A similar procedure could be used for sta
RNAse－A－Treatment－of－Mouse－Cells
IntroductionRNAse A treatment of permeabilized cells followed by immunostaining is a method which allows to show if the localization of a protein into
Mouse－Compete－Blood－Counts
Materials: 250 µL of fresh mouse blood in plastic tubes containing EDTA. RBC lysis buffer (388 mM NH4Cl, 29.7 mM NaHCO3, 25 µM Na2EDTA)20.75g. NH4CL2.
Shellless－cultures－of－chick－embryos
This experiment allows you to observe the development of a chick embryo outside its shell. Cultured chicks are also more accessible for manipulation.