Super‐resolution microscopy as a potential approach to diagnosis of platelet granule disorders

Autor: Daniel J. Metcalf, Keith Gomez, Michael Shaw, Jemima J. Burden, David Westmoreland, Daniel F. Cutler, William Grimes, Alex E. Knight
Jazyk: angličtina
Rok vydání: 2016
Předmět:
0301 basic medicine
Blood Platelets
Pathology
medicine.medical_specialty
Heterozygote
Genotype
Platelet Function Tests
diagnostic imaging
Platelet disorder
structured illumination microscopy
Hemorrhage
Cytoplasmic Granules
Antibodies
03 medical and health sciences
super‐resolution
Microscopy
medicine
Humans
Platelet
Frameshift Mutation
Blood Platelet Disorders
Hermansky‐Pudlak syndrome
CD63
Super-resolution microscopy
business.industry
Nucleotides
Platelet-Rich Plasma
Tetraspanin 30
Hematology
Original Articles
medicine.disease
PLATELETS
3. Good health
Microscopy
Electron
030104 developmental biology
Phenotype
Albinism
Oculocutaneous
Hermanski-Pudlak Syndrome
Platelet-rich plasma
Codon
Terminator
Original Article
platelet storage pool disorder
Hermansky–Pudlak syndrome
business
Gene Deletion
Zdroj: Journal of Thrombosis and Haemostasis
ISSN: 1538-7836
1538-7933
Popis: Essentials Deficiencies in size, number or shape of platelet granules are associated with bleeding symptoms. Super-resolution microscopy (SRM) facilitates the diagnosis of structural platelet disorders. SRM can deliver quantitative, automated, unbiased high-throughput morphometric analyses. Using CD63 as a marker, Hermansky-Pudlak patients are easily distinguished from controls. SummaryBackground Many platelet functions are dependent on bioactive molecules released from their granules. Deficiencies of these granules in number, shape or content are associated with bleeding. The small size of these granules is such that imaging them for diagnosis has traditionally required electron microscopy. However, recently developed super-resolution microscopes provide sufficient spatial resolution to effectively image platelet granules. When combined with automated image analysis, these methods provide a quantitative, unbiased, rapidly acquired dataset that can readily and reliably reveal differences in platelet granules between individuals. Objective To demonstrate the ability of structured illumination microscopy (SIM) to efficiently differentiate between healthy volunteers and three patients with Hermansky-Pudlak syndrome. Methods Blood samples were taken from three patients with Hermansky-Pudlak syndrome and seven controls. Patients 1–3 have gene defects in HPS1, HPS6 and HPS5, respectively; all controls were healthy volunteers. Platelet-rich plasma was isolated from blood and the platelets fixed, stained for CD63 and processed for analysis by immunofluorescence microscopy, using a custom-built SIM microscope. Results SIM can successfully resolve CD63-positive structures in fixed platelets. A determination of the number of CD63-positive structures per platelet allowed us to conclude that each patient was significantly different from all of the controls with 99% confidence. Conclusions A super-resolution imaging approach is effective and rapid in objectively differentiating between patients with a platelet bleeding disorder and healthy volunteers. CD63 is a useful marker for predicting Hermansky-Pudlak syndrome and could be used in the diagnosis of patients suspected of other platelet granule disorders.
Databáze: OpenAIRE
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