He has completed fellowship training in both intensive care medicine and emergency medicine, as well as post-graduate training in biochemistry, clinical toxicology, clinical epidemiology, and health professional education. He is an internationally recognised Clinician Educator with a passion for helping clinicians learn and for improving the clinical performance of individuals and collectives.Īfter finishing his medical degree at the University of Auckland, he continued post-graduate training in New Zealand as well as Australia’s Northern Territory, Perth and Melbourne. He is on the Board of Directors for the Intensive Care Foundation and is a First Part Examiner for the College of Intensive Care Medicine. He is also a Clinical Adjunct Associate Professor at Monash University. He is a co-founder of the Australia and New Zealand Clinician Educator Network (ANZCEN) and is the Lead for the ANZCEN Clinician Educator Incubator programme. Anaerobes: Consider brain abscess, elderlyĬhris is an Intensivist and ECMO specialist at the Alfred ICU in Melbourne.H Influenzae: (3%) – Head trauma with CSF leak, otitis, sinusitis, anatomical defects such as dermal sinus tracts.Staphylococcus: Penetrating skull injury, ear or neuro operations.N meningitidis: (30%) – Children and adolescents.Pneumococcus: (40%) – Otitis media, head injury, pneumonia, immunocompromised.Lymphocytosis, variable protein elevation and normal glucose.Aseptic meningitis (Generally accepted as mainly viral meningitis).Sickle cell disease – Capsulated organisms.Humoral or asplenic – Neiserria, enterovirus.N meningitidis, s. pneumonia, listeria, klebsiella, s. aureus.extended culture (Listeria, Cryptococcus).xanthochromic index with spectrophotometry (in SAH).less than half serum in infections (bacterial, Tb and fungal infections) and vasculitis and sarcoidosis.increased in CNS inflammation (including CSF drains and blood in CSF).oligoclonal bands in multiple sclerosis.increased in GBS, vasculitis and sarcoidosis.increased in infection: Tb > bacterial > viral.mixed lymphocytosis/monocytosis in GBS and status epilepticus.lymphocytosis: viral, TB, cryptococcal and listerial infections.polymorphonuclear leukocytosis: bacterial infection.in traumatic tap classically taught to expect 1 WCC : 500 RCC (if normal in peripheral cell counts) but this is not reliable.yellow with xanthochromia (this takes 6-12 hours to develop after blood enters CSF).blood stained with SAH and traumatic taps.cryptoccal antigen and Indian ink stain.Note that samples should not remain in a fixation buffer for extended periods of time as this can affect fluor conformation and fluorescence. The samples should be resuspended in Cell Staining Buffer. Note: If you are unable to immediately read your samples on a cytometer, keep them shielded from light and in a refrigerator set at 4-8☌. Perform fluorescence activated cell sorting (FACS), or flow cytometric analysis.Tip: For gentler fixation (particularly with tandem fluors), FluoroFix™ Buffer (Cat. Resuspend cells in 0.5ml Cell Staining Buffer or 0.5ml 2% paraformaldehyde-PBS fixation buffer.405204) and incubate for 15-20 minutes in the dark. If using a biotinylated primary antibody, resuspend cell pellet in residual buffer and add a previously determined optimum concentration of fluorochrome conjugated Streptavidin (SAv) reagent (e.g.FITC anti-mouse Ig) and incubate in the dark for 15-20 minutes. If using a purified primary antibody, resuspend pellet in residual buffer and add a previously determined optimum concentration of anti-species immunoglobulin fluorochrome conjugated secondary antibody (e.g.Wash 1X with at least 2ml of Cell Staining Buffer by centrifugation at 350xg for 5 minutes.Centrifuge at 350xg for 5 minutes, discard the supernatant.Incubate at room temperature for 10 minutes. Add 2ml of 1X RBC lysis solution to whole blood/antibody mixture. 420301) to 1X working concentration with DI water. Dilute 10X Red Blood Cell (RBC) Lysis Buffer (BioLegend Cat.Incubate at room temperature for 15-20 minutes in the dark.Add predetermined optimum concentrations of desired fluorochrome conjugated, biotinylated, or purified primary antibodies to 100µl of anti-coagulated whole blood.(Optional): Prior to staining with antibody of interest, add 5 µl of Human TruStain FcX™ per million cells in 100 µl staining volume, mix and incubate at room temperature for 5-10 minutes.TruStain FcX™ PLUS (anti-CD16/32, recommended for mouse cells.Human TruStain FcX™ (Fc Receptor Blocking Solution, BioLegend Cat.TruStain FcX™ (anti-CD16/32, BioLegend Cat.7-AAD Viability Staining Solution (BioLegend Cat.Cell Surface Flow Cytometry Staining of Whole Blood
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |