I have a LIVE/DEAD BacLight Bacterial Viability kit that has SYTO 9 and propidium iodide in it. Will I be able to stain eukaryotic cells that have engulfed bacteria and determine if the bacteria are alive or dead using this kit?
Unfortunately, no. SYTO 9 will label the nuclei of live or dead cells, including the eukaryotic cells. Propidium iodide is cell impermeant, and will only enter dead cells. If the eukaryotic cells are dead, they will label with propidium iodide as well. If the eukaryotic cells are alive, propidium iodide will not be able to enter and thus will not label the bacteria inside, whether the bacteria are alive or dead. We are not aware of any way to do a viability assay of bacteria once they have been engulfed by cells.
Is propidium iodide (PI) fixable with glutaraldehyde or paraformaldehyde (PFA)?
PI is not fixable with glutaraldehyde or PFA. Both reagents fix by crosslinking amines. PI and other nucleic acid stains do not inherently bind covalently to nucleic acids and these fixatives do not crosslink the dyes to nucleic acids.
The one fixable nucleic acid stain is Ethidium Monoazide Bromide (EMA), Cat no. E1374); it covalently binds to nucleic acids upon activation by exposure to light.
Signaling through MHC class II molecules blocks CD95-induced apoptosis.
Authors:Catlett IM,Xie P,Hostager BS,Bishop GA
Journal:Journal of immunology (Baltimore, Md. : 1950)
PubMed ID:11342618
Glutamate and non-glutamate receptor mediated toxicity caused by oxygen and glucose deprivation in organotypic hippocampal cultures.
Authors:Newell DW, Barth A, Papermaster V, Malouf AT
Journal:J Neurosci
PubMed ID:7472521
In vitro ischemia models have utilized oxygen, or oxygen and glucose deprivation to simulate ischemic neuronal injury. Combined oxygen and glucose deprivation can induce neuronal damage which is in part mediated through NMDA receptors. Severe oxygen deprivation alone however can cause neuronal injury which is not NMDA mediated. We tested ... More
Identification and characterization of two subpopulations of Encephalitozoon intestinalis.
Authors:Hoffman RM, Marshall MM, Polchert DM, Jost BH
Journal:Appl Environ Microbiol
PubMed ID:12902292
Microsporidia are obligate intracellular protozoa that have been shown to be pathogenic to most living creatures. The development of in vitro cell culture propagation methods has provided researchers with large numbers of spores and facilitated the study of these organisms. Here, we describe heterogeneity within cell culture-propagated Encephalitozoon intestinalis suspensions. ... More
Autoantigens targeted in systemic lupus erythematosus are clustered in two populations of surface structures on apoptotic keratinocytes.
Authors:Casciola-Rosen LA, Anhalt G, Rosen A
Journal:J Exp Med
PubMed ID:7511686
'Systemic lupus erythematosus is a multisystem autoimmune disease in which the autoantibody response targets a variety of autoantigens of diverse subcellular location. We show here that these autoantigens are clustered in two distinct populations of blebs at the surface of apoptotic cells. The population of smaller blebs contains fragmented endoplasmic ... More
Caspase activation contributes to delayed death of heat-stressed striatal neurons.
Authors:White MG, Emery M, Nonner D, Barrett JN
Journal:J Neurochem
PubMed ID:14622126
'Hyperthermia can contribute to brain damage both during development and post-natally. We used rat embryonic striatal neurons in culture to study mechanisms underlying hyperthermia-induced neuronal death. Heat stress at 43 degrees C for 2 h produced no obvious signs of damage during the first 12 h after the stress, but ... More