Cytological Effects of Prefixation Treatment

Hungerford, David A.; DiBerardino, Marie A.

doi:10.1083/jcb.4.4.391

Cited by 37 publications

(2 citation statements)

References 13 publications

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“…This conservation of the characteristic structure of chromosomes in solutions containing 100 µM K-Hepes buffer as the only ionic component contrasted with the large increase in volume of chromosomes isolated by conventional procedures [30] – [32] and of chromosomes in situ [33] , [34] in media of low ionic strength. To confirm that their structure was not influenced by contaminating cations in the solutions of crowding macromolecules, these were assayed by atomic emission spectrometry.…”

Section: Resultsmentioning

confidence: 99%

Structure of Metaphase Chromosomes: A Role for Effects of Macromolecular Crowding

Hancock

2012

PLoS ONE

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In metaphase chromosomes, chromatin is compacted to a concentration of several hundred mg/ml by mechanisms which remain elusive. Effects mediated by the ionic environment are considered most frequently because mono- and di-valent cations cause polynucleosome chains to form compact ∼30-nm diameter fibres in vitro, but this conformation is not detected in chromosomes in situ. A further unconsidered factor is predicted to influence the compaction of chromosomes, namely the forces which arise from crowding by macromolecules in the surrounding cytoplasm whose measured concentration is 100–200 mg/ml. To mimic these conditions, chromosomes were released from mitotic CHO cells in solutions containing an inert volume-occupying macromolecule (8 kDa polyethylene glycol, 10.5 kDa dextran, or 70 kDa Ficoll) in 100 µM K-Hepes buffer, with contaminating cations at only low micromolar concentrations. Optical and electron microscopy showed that these chromosomes conserved their characteristic structure and compaction, and their volume varied inversely with the concentration of a crowding macromolecule. They showed a canonical nucleosomal structure and contained the characteristic proteins topoisomerase IIα and the condensin subunit SMC2. These observations, together with evidence that the cytoplasm is crowded in vivo, suggest that macromolecular crowding effects should be considered a significant and perhaps major factor in compacting chromosomes. This model may explain why ∼30-nm fibres characteristic of cation-mediated compaction are not seen in chromosomes in situ. Considering that crowding by cytoplasmic macromolecules maintains the compaction of bacterial chromosomes and has been proposed to form the liquid crystalline chromosomes of dinoflagellates, a crowded environment may be an essential characteristic of all genomes.

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Section: Resultsmentioning

confidence: 99%

Structure of Metaphase Chromosomes: A Role for Effects of Macromolecular Crowding

Hancock

2012

PLoS ONE

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“…Hungerford had experimented with various combinations of prefixation treatment with salt solutions (25, 26) and exposure to colchicine (27) reported by others and made chromosome preparations by the squash technique (28). With Marie DiBerardino, he made a detailed investigation of the effects of various pre-treatment protocols on cells of mouse and frog embryos (29). The experience gained from these time-consuming investigations would prove useful in his subsequent work with Nowell on chromosomes of leukemic cells.…”

Section: Reminiscencesmentioning

confidence: 99%

Philadelphia Chromosome Symposium: commemoration of the 50th anniversary of the discovery of the Ph chromosome

Chandra¹,

Heisterkamp²,

Hungerford³

et al. 2011

Cancer Genetics

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This report summarizes highlights of the ‘Philadelphia Chromosome Symposium: Past, Present and Future’, held September 28, 2010, to commemorate the 50th anniversary of the discovery of the Philadelphia chromosome. The symposium sessions included presentations by investigators who made seminal contributions concerning the discovery and molecular characterization of the Ph chromosome and others who developed a highly successful therapy based on the specific molecular alteration observed in chronic myelogenous leukemia. Additional presentations highlighted future opportunities for the design of molecularly targeted therapies for various types of cancer. Also included here are reminiscences connected with the discovery of the Ph chromosome by David Hungerford and Peter Nowell, the discovery that the abnormality arises from a chromosomal translocation, by Janet Rowley, and the cloning of the 9;22 translocation breakpoints by Nora Heisterkamp, John Groffen and colleagues.

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Validation of a novel alcohol‐free preservative solution in comparison with conventional prefixation on quality of serous body fluid smears

Ambalaparambil,

Perumal,

Gochhait

et al. 2024

Diagnostic Cytopathology

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BackgroundWe describe a novel alcohol‐free preservative composed of glucose, mannitol, disodium hydrogen orthophosphate, thymol, and distilled water (glucose‐mannitol–disodium dihydrogen orhtophosphate‐thymol [GMDT] preservative) in appropriate proportion as an alternative to alcohol prefixation (APF) of body fluids.ObjectivesTo assess the cytomorphologic preservation and staining quality of serous body fluid smears generated by GMDT preservative and compare it with smears processed by standard 50% APF.MethodologyThe study comprised 151 effusion samples. Each sample was equally divided into four tubes. Equal volumes of APF and GMDT preservatives were added to the first two tubes and left at room temperature for 24 h. Similarly, the corresponding preservatives were added to the third and fourth tubes and stored for 48 h. Two smears were prepared from the centrifuged sediments of each tube (all four tubes) and stained with May‐Grünwald Giemsa and Papanicolaou (Pap) stains. Using a three‐tiered scoring system, the smear examination was blinded to assess the extent of cellular preservation and the staining quality by two cytotechnologists and two cytopathologists. Statistical analysis was performed by STATA 16.0.ResultsSamples processed with the GMDT preservative at 24 h showed better cytoplasmic preservation and smear background, while nuclear features and staining quality showed no difference between the two preservatives. Mild cytoplasmic and nuclear degenerative changes were noted with the GMDT at 48 h, while all four parameters remained similar with APF at 24 and 48 h.ConclusionsThe newly developed alcohol‐free, GMDT preservative, could be a feasible and cost‐effective alternative to 50% APF, preferably when samples are processed within 24 h.

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Cytological Effects of Prefixation Treatment

Cited by 37 publications

References 13 publications

Structure of Metaphase Chromosomes: A Role for Effects of Macromolecular Crowding

Structure of Metaphase Chromosomes: A Role for Effects of Macromolecular Crowding

Philadelphia Chromosome Symposium: commemoration of the 50th anniversary of the discovery of the Ph chromosome

Validation of a novel alcohol‐free preservative solution in comparison with conventional prefixation on quality of serous body fluid smears

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