Differences In Cell Size Research Articles

Double flower formation induced by silencing of C-class MADS-box genes and its variation among petunia Cultivars

Double flowers induced by virus-induced gene silencing (VIGS) of two C-class MADS-box genes, pMADS3 and FBP6, were investigated in four cultivars of Petunia hybrida. In flowers induced by either pMADS3-VIGS or FBP6-VIGS, only small changes in commercial appearance were recognized regardless of cultivar, whereas in those induced by pMADS3/FBP6-VIGS, complete conversion of stamens into petaloid tissues and marked enlargement of upper limb-like tissues were observed, resulting in a decorative appearance in all the four cultivars. In whorl 4, cultivar-dependent conversion of carpels into new flowers was noted in pMADS3/FBP6-VIGS flowers. Of the four cultivars, only ‘Mambo Purple’ exhibited the development of new flowers instead of carpels and the emergence of ectopic new flowers from the axil of whorl 3 organs, which created an ornamental appearance with a high commercial value. Investigation of large and small petaloid stamens induced by pMADS3/FBP6-VIGS and pMADS3-VIGS, respectively, revealed only small differences in cell size compared to the large difference in total surface area. Quantitative RT-PCR analysis of SQUAMOSA/AP1/FRU type A-class genes, FBP29, PFG, and FBP26, showed no close relationship between the expression of those genes and petaloid stamen size. The suppressed C-class gene function at the late stage of flower development has little influence on the final size of petaloid tissue.

Adipose tissue and metabolic alterations: regional differences in fat cell size and number matter, but differently: a cross-sectional study.

White adipose tissue can expand by increasing the size and/or number of fat cells. Although increased sc and visceral fat cell size associates with an adverse metabolic profile, the relationship with fat cell number in either depot is unknown. We hypothesized that adipocyte number and size displayed different relationships with clinically relevant metabolic variables. This was a cross-sectional study of 204 patients scheduled for gastric bypass surgery. Fat cell size and number were determined in visceral and abdominal sc adipose tissue and related to insulin sensitivity (by hyperinsulinemic euglycemic clamp), fasting plasma levels of insulin, triglycerides and high-density lipoprotein (HDL) cholesterol. Visceral and sc fat cell volumes were positively correlated with insulin and triglyceride levels and negatively with insulin sensitivity and HDL-cholesterol (P = .0020 or better). In contrast, although visceral fat cell number did not associate with any metabolic parameter, sc adipocyte number displayed a positive association with insulin sensitivity and HDL-cholesterol and a negative relationship with insulin and triglyceride levels (P = .0014 or better). All results were independent of body fat mass. Variations in fat cell size and number correlate differently with metabolic parameters in obesity. Increased fat cell size in visceral and sc depots associates with a pernicious metabolic profile, whereas increased sc, but not visceral, fat cell number correlates with a more beneficial phenotype. Whether determination of sc fat cell number, in addition to adipocyte size, may have a predictive value for the risk of type 2 diabetes needs to be demonstrated in prospective or mechanistic studies.

Differences in benthic diatom assemblages between streams and lakes in Sweden and implications for ecological assessment

Benthic diatom assemblages from 179 streams and 98 lakes in Sweden were compared to identify differences in taxonomic composition, growth form, cell size, diversity and richness, and ultimately, to assess the effects of differences between streams and lakes on assessments. Streams and lakes with similar environmental background variables were chosen to avoid an overriding effect of factors, such as nutrients and pH, on taxonomic composition. Generic and species composition differed significantly between streams and lakes. Richness was significantly higher in lakes than in streams, and the average diatom cell was significantly longer in streams than in lakes. No other measures analyzed differed significantly. Diatom assemblages in both habitats were related to pH and nutrients, and the diatom indices studied responded similarly to the main environmental gradients in both habitats. Thus, despite differences in taxonomic composition between streams and lakes, the resulting diatom assemblages are similar, enabling use of the same diatom indices in both habitats.

A system for counting fetal and maternal red blood cells.

The Kleihauer-Betke (KB) test is the standard method for quantitating fetal-maternal hemorrhage in maternal care. In hospitals, the KB test is performed by a certified technologist to count a minimum of 2000 fetal and maternal red blood cells (RBCs) on a blood smear. Manual counting suffers from inherent inconsistency and unreliability. This paper describes a system for automated counting and distinguishing fetal and maternal RBCs on clinical KB slides. A custom-adapted hardware platform is used for KB slide scanning and image capturing. Spatial-color pixel classification with spectral clustering is proposed to separate overlapping cells. Optimal clustering number and total cell number are obtained through maximizing cluster validity index. To accurately identify fetal RBCs from maternal RBCs, multiple features including cell size, roundness, gradient, and saturation difference between cell and whole slide are used in supervised learning to generate feature vectors, to tackle cell color, shape, and contrast variations across clinical KB slides. The results show that the automated system is capable of completing the counting of over 60,000 cells (versus ∼2000 by technologists) within 5 min (versus ∼15 min by technologists). The throughput is improved by approximately 90 times compared to manual reading by technologists. The counting results are highly accurate and correlate strongly with those from benchmarking flow cytometry measurement.

Enzymatic modification and particle size reduction of wheat bran improves the mechanical properties and structure of bran-enriched expanded extrudates

The aim of this study was to examine enzymatic modification of wheat bran, performed in a low-moisture process, and the reduction of bran particle size as means of improving the technological performance of wheat bran in expanded extrudates. Modification of bran by hydrolytic enzymes increased the crispiness and decreased the hardness and piece density of extrudates containing wheat bran and endosperm rye flour in 20:80 ratio. These improvements correlated (P < 0.01 or 0.05) with an increased content of water extractable arabinoxylan and decreased water holding capacity of the bran, as well as with increased longitudinal expansion of the extrudates. Furthermore, bran with a fine average particle size (84 μm) produced extrudates with improved mechanical properties and higher radial expansion than coarse bran (particle size 702 μm). The impact of bran particle size was also observed in the cellular structure of the extrudates as differences in cell size and homogeneity. The bran drying method, oven or freeze drying after enzymatic modification, did not have a major impact on the properties of the extrudates. The study showed that the functionality of wheat bran in extrusion can be improved by enzymatic modification using a low-water process and by reduction of bran particle size.

Comparison of Morphology of Bulbar Conjunctival Cells Assessed by Impression Cytology versus Scrape and Smear Methods

Purpose: To assess the morphology of superficial bulbar conjunctival cells collected by impression cytology or by scraping.Methods: Freshly procured sheep eyes from a slaughterhouse were washed and then conjunctival impression cytology (CIC) samples taken from the nasal aspect of the bulbar conjunctiva on a Biopore Millicell filter unit, or cells scraped from the same location and smeared onto glass slides. After alcohol fixation, all samples were stained with Giemsa. Cell images were obtained by light microscopy at 200× magnification, and subjected to morphometry.Results: Cells obtained by scrape/smear processing appeared larger and less densely staining than cells procured by CIC. This appearance was confirmed by morphometry with measures of cell longest dimension LONG being 27.7 ± 1.3 μm and 21.7 ± 1.5 μm, respectively, while cell AREAC measures averaged 543 ± 42 μm2 and 333 ± 51 μm2; these differences were significant (p < 0.001). Similarly, cell nucleus length (NUCLONG) and nucleus area (AREANUC) were also larger in smeared cells (p < 0.001). Various calculations of nucleo-cytoplasmic ratios as based on dimensions or area were only slightly different when comparing cells obtained by the two methods, indicating that the glass slide-smeared cells simply spread out more than cells on the CIC filters.Conclusions: Differences in cell and nucleus size can result from use of different processing methods. Therefore, in making comparisons between sets of conjunctival cells, it is important that cell collection and processing methods be considered.

Production of CMAH Knockout Preimplantation Embryos Derived From Immortalized Porcine Cells Via TALE Nucleases

Although noncancerous immortalized cell lines have been developed by introducing genes into human and murine somatic cells, such cell lines have not been available in large domesticated animals like pigs. For immortalizing porcine cells, primary porcine fetal fibroblasts were isolated and cultured using the human telomerase reverse transcriptase (hTERT) gene. After selecting cells with neomycin for 2 weeks, outgrowing colonized cells were picked up and subcultured for expansion. Immortalized cells were cultured for more than 9 months without changing their doubling time (~24 hours) or their diameter (< 20 µm) while control cells became replicatively senescent during the same period. Even a single cell expanded to confluence in 100 mm dishes. Furthermore, to knockout the CMAH gene, designed plasmids encoding a transcription activator-like effector nuclease (TALENs) pairs were transfected into the immortalized cells. Each single colony was analyzed by the mutation-sensitive T7 endonuclease I assay, fluorescent PCR, and dideoxy sequencing to obtain three independent clonal populations of cells that contained biallelic modifications. One CMAH knockout clone was chosen and used for somatic cell nuclear transfer. Cloned embryos developed to the blastocyst stage. In conclusion, we demonstrated that immortalized porcine fibroblasts were successfully established using the human hTERT gene, and the TALENs enabled biallelic gene disruptions in these immortalized cells.

Subcutaneous adipose cell size and distribution: Relationship to insulin resistance and body fat

Metabolic heterogeneity among obese individuals may be attributable to differences in adipose cell size. We sought to clarify this by quantifying adipose cell-size distribution, body fat, and insulin-mediated glucose uptake in overweight/moderately-obese individuals. 148 healthy nondiabetic subjects with BMI 25–38 kg/m2 underwent subcutaneous adipose tissue biopsies and quantification of insulin-mediated glucose uptake with steady-state plasma glucose concentrations (SSPG) during the modified insulin suppression test. Cell-size distributions were obtained with Beckman Coulter Multisizer. Primary endpoints included % small adipose cells and diameter of large adipose cells. Cell-size and metabolic parameters were compared by regression for the whole group; according to IR and IS subgroups; and by body fat quintile.Both large and small adipose cells were present in nearly equal proportions. Percent small cells was associated with SSPG (r=0.26, p=0.003). Compared to BMI-matched IS individuals, IR counterparts demonstrated fewer, but larger large adipose cells, and a greater proportion of small-to-large adipose cells. Diameter of the large adipose cells was associated with %body fat (r=0.26, p=0.014), female sex (r=0.21, p=0.036), and SSPG (r=0.20, p=0.012). In the highest vs lowest % body fat quintile, adipose cell size increased by only 7% whereas adipose cell number increased by 74%.Recruitment of adipose cells is required for expansion of body fat mass beyond BMI of 25 kg/m2. Insulin resistance is associated with accumulation of small adipose cells and enlargement of large adipose cells. These data support the notion that impaired adipogenesis may underlie insulin resistance.

Morphology and size of stem cells from mouse and whale: observational study

Objective To compare the morphology and size of stem cells from two mammals of noticeably different body size.Design Observational study.Setting The Netherlands.Participants A humpback whale (Megaptera novaeangliae) and a laboratory...

Adipocyte Mitochondrial Function Is Reduced in Human Obesity Independent of Fat Cell Size

It has been suggested that mitochondrial dysfunction in adipocytes contributes to obesity-related metabolic complications. However, obesity results in adipocyte hypertrophy, and large and small adipocytes from the same depot have different characteristics, raising the possibility that obesity-related mitochondrial defects are an inherent function of large adipocytes. Our goal was to examine whether obesity, independent of fat cell size and fat depot, is associated with mitochondria dysfunction. We compared adipocyte mitochondrial function using a cross-sectional comparison study design. The studies were performed at Mayo Clinic, an academic medical center. Omental and/or abdominal subcutaneous adipose samples were collected from 20 age-matched obese and nonobese nondiabetic men and women undergoing either elective abdominal surgery or research needle biopsy. Interventions were not conducted as part of these studies. We measured mitochondrial DNA abundance, oxygen consumption rates, and citrate synthase activity from populations of large and small adipocytes (separated with differential floatation). For both omental and subcutaneous adipocytes, at the cell and organelle level, oxygen consumption rates and citrate synthase activity were significantly reduced in cells from obese compared with nonobese volunteers, even when matched for cell size by comparing large adipocytes from nonobese and small adipocytes from obese. Adipocyte mitochondrial content was not significantly different between obese and nonobese volunteers. Mitochondrial function and content parameters were not different between small and large cells, omental, and subcutaneous adipocytes from the same person. Adipocyte mitochondrial oxidative capacity is reduced in obese compared with nonobese adults and this difference is not due to cell size differences. Adipocyte mitochondrial dysfunction in obesity is therefore related to overall adiposity rather than adipocyte hypertrophy.

Adipose tissue depot and cell size dependency of adiponectin synthesis and secretion in human obesity

Adiponectin is an insulin sensitizing fat cell (FC) hormone whose levels are related to adipose tissue (AT) mass and depot distribution. We hypothesized that the nature of AT expansion (hypertrophy vs. hyperplasia) contributes to obesity-related reductions in serum adiponectin and that this effect is influenced by the regional distribution of AT to subcutaneous (S) and visceral (V) depots. Thirteen obese subjects provided paired AT biopsies. Serum total and high molecular weight (HMW) adiponectin levels were determined by ELISA. Secretion was quantified following 24-h explant culture. FC size, number, % large, and % small FC were determined by microscopic analysis. Secretion of total adiponectin was highest by SAT (P = 0.008) and correlated more strongly with serum adiponectin (total: P = 0.015, r = 0.77; HMW: P = 0.005, r = 0.83) than did secretion by VAT (P = 0.05, r = 0.66 for both). FC size was greatest in SAT and correlated negatively with both serum (total: P = 0.01, r = −0.74; HMW: P = 0.03, r = −0.69) and secreted (total: P = 0.05, r = −0.72; HMW: P = 0.02, r = −0.87) adiponectin. The % small FC in SAT correlated positively with both serum (total: P = 0.006, r = 0.87; HMW: P = 0.009, r = 0.79) and secreted (total: P = 0.03, r = 0.75; HMW: P = 0.01, r = 0.92) adiponectin. VAT FC size correlated negatively with serum HMW adiponectin (P = 0.01, r = −0.76) but not with any measure of secretion. VAT had the greatest % small FC, which related positively to serum HMW (P = 0.004, r = 0.81) and to secreted total adiponectin (P = 0.02, r = 0.78). These studies indicate that differences in fat cell size and depot distribution of AT expansion are important influences on adiponectin in obesity.

Optimizing the use of rainbow trout hepatocytes for bioaccumulation assessments with fish

Biotransformation rates measured using cryopreserved trout hepatocytes can be extrapolated to the whole animal to predict metabolism impacts on chemical bioaccumulation. Future use of these methods within a regulatory context requires, however, that they be optimized and standardized. Specifically, questions exist concerning gender differences in metabolism, cryopreservability of cells, and the accuracy of in vitro–in vivo scaling factors.2. In this study, we evaluated hepatocytes from juvenile male and female trout. No gender differences in cell size, protein abundance, cytochrome P450 content, ethoxyresorufin-O-deethylase activity, uridine diphosphate glucuronosyltransferase activity or intrinsic clearance of pyrene were observed for freshly isolated hepatocytes. There was a small difference in measured glutathione-S-transferase activity (<25%; males > females).3. Cells were cryopreserved by two methods: direct placement into liquid N2 vapor and controlled, slow-rate freezing. Comparable live recovery and enzymatic activity were observed regardless of freezing method or gender. Cells cryopreserved in liquid N2 vapor exhibited activity levels similar to those of freshly isolated cells, although there were small but significant differences in pyrene clearance and glutathione-S-transferase activity (frozen < fresh). Hepatocellularity values did not differ by sex.4. These results suggest that hepatocytes from male and female juvenile trout may be used interchangeably for in vitro–in vivo metabolism extrapolations.

Comparative Proteomics Reveals a Role for Seed Storage Protein AmA1 in Cellular Growth, Development, and Nutrient Accumulation

Seed storage proteins are known to be utilized as carbon and nitrogen source for growing seedlings and thus are considered as potential candidates for nutritional improvement. However, their precise function remains unknown. We have earlier shown that ectopic expression of a seed storage protein, AmA1, leads to increase in protein besides high tuber yield in potato. To elucidate the AmA1-regulated molecular mechanism affecting increased protein synthesis, reserve accumulation, and enhanced growth, a comparative proteomics approach has been applied to tuber life-cycle between wild-type and AmA1 potato. The differential display of proteomes revealed 150 AmA1-responsive protein spots (ARPs) that change their intensities more than 2.5-fold. The LC-ESI-MS/MS analyses led to the identification of 80 ARPs presumably associated with cell differentiation, regulating diverse functions, viz., protein biogenesis and storage, bioenergy and metabolism, and cell signaling. Metabolome study indicated up-regulation of amino acids paralleling the proteomics analysis. To validate this, we focused our attention on anatomical study that showed differences in cell size in the cortex, premedullary zone and pith of the tuber, coinciding with AmA1 expression and localization. Further, we interrogated the proteome data using one-way analysis of variance, cluster, and partial correlation analysis that identified two significant protein modules and six small correlation groups centered around isoforms of cysteine protease inhibitor, actin, heat shock cognate protein 83 and 14-3-3, pointing toward AmA1-regulated overlapping processes of protein enhancement and cell growth perhaps through a common mechanism of function. A model network was constructed using the protein data sets, which aim to show how target proteins might work in coordinated fashion and attribute to increased protein synthesis and storage reserve accumulation in AmA1 tubers on one hand and organ development on the other.

Settling and bioflocculation of two species of algae used in wastewater treatment and algae biomass production

The settling and bioflocculation of two strains of algae were investigated in the laboratory to provide insights to help improve algae settling in large‐scale, algae‐based wastewater treatment systems with simultaneous algae biofuel production. Energy‐efficient algae harvesting methods are needed for cost‐competitive production of biofuels from algae. The use of bacterially derived exudates (bioflocculation) is promising, but its use in high‐rate algae pond (HRAP) wastewater systems has not yielded consistently reliable settling. The settling of the algae species Scenedesmus sp. and Chlorella vulgaris was investigated in the laboratory with and without cultures of the bacterium Burkholderia cepacia to investigate environmental effects on the settling of pure algal cultures and to examine possible bioflocculation by bacteria and bacterial exudates. Scenedesmus sp. and C. vulgaris responded in opposite ways to changes in operating conditions such as culture age and bioflocculation. Scenedesmus sp. settled better in later stationary growth stages, while C. vulgaris settled better in its early exponential growth phase. Scenedesmus sp. settling was improved by bioflocculation during early exponential and late stationary growth stages, but not during early stationary growth. Scenedesmus sp. responded better to B. cepacia cells plus filtrate (filtered bacterial broth) than to filtrate addition alone. In contrast, C. vulgaris settling was not improved via bioflocculation with neither the addition of B. cepacia cells nor filtrate at any growth stage. Longer contact time between the algae cells and bioflocculants improved the settling of Scenedesmus sp., but not the settling of C. vulgaris. Observed differences in settling and bioflocculation of the two algae species were attributed to differences in algal cell size and shape, culture density, and exudate type (i.e., capsular versus dissolved EPS). The widely different settling behavior of the two species of algae used in this research may explain the often varied results observed with mixed cultures in algae wastewater ponds. © 2013 American Institute of Chemical Engineers Environ Prog, 32: 946–954, 2013

Proteomic and Comparative Genomic Analysis of Two Brassica napus Lines Differing in Oil Content

Ultrastructural observations, combined with proteomic and comparative genomic analyses, were applied to interpret the differences in protein composition and oil-body characteristics of mature seed of two Brassica napus lines with high and low oil contents of 55.19% and 36.49%, respectively. The results showed that oil bodies were arranged much closer in the high than in the low oil content line, and differences in cell size and thickness of cell walls were also observed. There were 119 and 32 differentially expressed proteins (DEPs) of total and oil-body proteins identified. The 119 DEPs of total protein were mainly involved in the oil-related, dehydration-related, storage and defense/disease, and some of these may be related to oil formation. The DEPs involved with dehydration-related were both detected in total and oil-body proteins for high and low oil lines and may be correlated with the number and size of oil bodies in the different lines. Some genes that corresponded to DEPs were confirmed by quantitative trait loci (QTL) mapping analysis for oil content. The results revealed that some candidate genes deduced from DEPs were located in the confidence intervals of QTL for oil content. Finally, the function of one gene that coded storage protein was verified by using a collection of Arabidopsis lines that can conditionally express the full length cDNA from developing seeds of B. napus.

Growth, photosynthetic efficiency, and biochemical composition of Tetraselmis suecica F&amp;M‐M33 grown with LEDs of different colors

The effect of light quality on cell size and cell cycle, growth rate, productivity, photosynthetic efficiency and biomass composition of the marine prasinophyte Tetraselmis suecica F&M-M33 grown in 2-L flat panel photobioreactors illuminated with light emitting diodes (LEDs) of different colors was investigated. Biomass productivity and photosynthetic efficiency were comparable between white and red light, while under blue and green light productivity decreased to less than half and photosynthetic efficiency to about one third. Differences in cell size and number correlated with the cell cycle phase. Under red light cells were smaller and more motile. Chlorophyll content was strongly reduced with red and enhanced with blue light, while carotenoids and gross biomass composition were not affected by light quality. The eicosapentaenoic acid content increased under red light. Red light can substitute white light without affecting productivity of T. suecica F&M-M33, leading to smaller and more motile cells and increased eicosapentaenoic acid content. Red LEDs can thus be profitably used for the production of this microalga for aquaculture.

A biologically inspired hierarchical goal directed navigation model

We propose an extended version of our previous goal directed navigation model based on forward planning of trajectories in a network of head direction cells, persistent spiking cells, grid cells, and place cells. In our original work the animat incrementally creates a place cell map by random exploration of a novel environment. After the exploration phase, the animat decides on its next movement direction towards a goal by probing linear look-ahead trajectories in several candidate directions while stationary and picking the one activating place cells representing the goal location. In this work we present several improvements over our previous model. We improve the range of linear look-ahead probes significantly by imposing a hierarchical structure on the place cell map consistent with the experimental findings of differences in the firing field size and spacing of grid cells recorded at different positions along the dorsal to ventral axis of entorhinal cortex. The new model represents the environment at different scales by populations of simulated hippocampal place cells with different firing field sizes. Among other advantages this model allows simultaneous constant duration linear look-ahead probes at different scales while significantly extending each probe range. The extension of the linear look-ahead probe range while keeping its duration constant also limits the degrading effects of noise accumulation in the network. We show the extended model’s performance using an animat in a large open field environment.

Extrapulmonary neuroendocrine small and large cell carcinomas: a review of controversial diagnostic and therapeutic issues

Extrapulmonary neuroendocrine carcinoma (EPNEC) is a heterogeneous and rare group of high-grade neoplasms occurring in different organs. They usually share a poor prognosis, but diagnostic and therapeutic options still include several controversial issues, due to the rarity of this condition and to differences in architecture and cell size, being some cases pure small cell carcinomas, other pure large cell neuroendocrine carcinomas and some others combined/mixed neuroendocrine carcinomas with a conventional non-neuroendocrine carcinoma. In addition, the therapeutic strategy varies in different organs (surgery and/or chemotherapy and/or radiation therapy and/or targeted treatments), and clinicians and pathologists are asked to interact to reach an accurate classification of every single case, as well as the most appropriate selection of the treatment options, even considering different time points of each EPNEC natural history. This overview highlights controversial pathological and clinical issues and summarizes possible solutions to most of such EPNEC-related problems.

An analysis of the spectrum of the discontinuous Galerkin method II: Nonuniform grids

We analyze the eigenvalues of the discontinuous Galerkin spatial operator for the one-dimensional linear advection equation on nonuniform grids. We show that when the difference in cell sizes is below an order-dependent critical number, the spectrum continuously changes with changing cell sizes. In a particularly simple case where the mesh contains cells of only two sizes, the spectrum grows linearly with the proportion of smaller cells in the mesh. When the cell size scale is larger than a critical value, the eigenvalues corresponding to smaller cells separate from the rest of the spectrum. We provide an easily computable estimate of the upper bound on the spectrum for both cases. This estimate can be used to compute a relaxed time step restriction if smaller cells are dispersed among larger cells. Numerical examples for one- and two-dimensional problems reveal that computational saving can be realized by use of a larger stable time step.

FACS purification of Drosophila larval neuroblasts for next-generation sequencing

Elegant tools are available for the genetic analysis of neural stem cell lineages in Drosophila, but a methodology for purifying stem cells and their differentiated progeny for transcriptome analysis is currently missing. Previous attempts to overcome this problem either involved using RNA isolated from whole larval brain tissue or co-transcriptional in vivo mRNA tagging. As both methods have limited cell type specificity, we developed a protocol for the isolation of Drosophila neural stem cells (neuroblasts, NBs) and their differentiated sibling cells by FACS. We dissected larval brains from fly strains expressing GFP under the control of a NB lineage-specific GAL4 line. Upon dissociation, we made use of differences in GFP intensity and cell size to separate NBs and neurons. The resulting cell populations are over 98% pure and can readily be used for live imaging or gene expression analysis. Our method is optimized for neural stem cells, but it can also be applied to other Drosophila cell types. Primary cell suspensions and sorted cell populations can be obtained within 1 d; material for deep-sequencing library preparation can be obtained within 4 d.