Journal of Biotechnology & Microbiology-Juniper Publishers Abstract The CellROX® Deep Red flow cytometry kit (Life technologies) has been developed for the detection of oxidative stress in mammalian cells. It combines a ROS sensitive fluorophore (CellROX® Deep Red) and a "viability” dye (SYTOX® Blue) to allow the detection of non-oxidized, oxidized and damaged cells. The present study investigated the application of these markers to Enterococcus faecalis and Fusobacterium nucleatum subjected to oxidative stress. An optimal concentration of CellROX® has been determined on Enterococcus faecalis and Fusobacterium nucleatum exposed to oxidative stress (H2O2). Bacteria have been exposed to various H2O2 concentrations and labeled with CellROX® to verify that fluorescence increased along with oxidative stress. Also, bacteria exposed to H2O2 were double stained with CellROX® and SYTOX® Blue and analyzed by flow cytometry. The optimal concentration of CellROX® Deep Red was 4^M for both strains. Fluorescence of bacteria labeled with 4^M of CellROX® Deep Red increased accordingly with the oxidative stress applied. Flow cytometry analysis of double stained samples showed bacteria subpopulations with increased CellROX® signal when stressed, and higher SYTOX® Blue uptake under higher oxidative stress. Results indicate that CellROX® Deep Red can be applied to measure oxidative stress in E. faecalis and F. nucleatum. The combination of CellROX® and SYTOX® Blue allowed the discrimination of non-oxidized, oxidized and damaged bacteria. Keywords: Oxidative stress; ROS; Enterococcus faecalis; Fusobacterium nucleatum; CellROX™ Deep Red; Flow cytometry Go to Introduction Free radicals are defined as highly reactive chemicals exhibiting unpaired electrons such as reactive oxygen species (ROS) [1]. Accumulation of ROS in cells damages nucleic and amino acids, membrane lipids and can initiate self-propagating oxidative chain reactions [2-5]. Developing methods able to correlate oxidative stress with bacterial response and survival would help to gain insight into the development of new antibacterial strategies. Unfortunately, the propensity of ROS to acquire electrons renders them highly reactive, short lived, and therefore very difficult to detect [6,7]. The use of ROS sensitive probes, that can be detected using several analytical methods such as spectrofluorometry, fluorescence microscopy or flow cytometry, offer high sensitivity and experimental convenience [8-10]. The ROS reporters hydroxy-phenyl-fluorescein and "singlet oxygen sensor green" have been previously employed to detect hydroxyl radicals and singlet oxygen in a cell-free model [11]. Dichlorofluoresceindiacetate and flow cytometry (FCM) were used by Subramanian et al. [12] to evaluate whether resveratrol, a redox active phytoalexin, can induce oxidative stress in E. coli [12]. However, the combined use of a ROS reporter with a "viability” marker to simultaneously assess oxidative stress and viability has never been investigated in bacteria. The CellROX® Deep Red flow cytometry assay kit (Life technologies), recently developed to assess the effects of oxidative stress in mammalian cells, comprises two dyes, namely: the CellROX® Deep Red, a ROS reporter, and the SYTOX® Blue, a "viability” dye. CellROX® Deep Red is cell permeable, cytoplasmic and does not fluoresce in a reduced state, while it becomes fluorescent upon oxidation. The second dye, SYTOX® Blue, is a cyanine nucleic acid stain which only diffuses into membrane-damaged cells. Only one epifluorescence microscopy study has used the CellROX® Deep Red to visualize oxidative stress in ionophore-treated Bacillus subtilis, whereas SYTOX® Blue has been previously used to assess bacterial membrane integrity by flow cytometry [13-15]. However, flow cytometry (FCM) has never been applied to correlate oxidative stress and viability in bacteria using the combination of these two dyes. The aim of this study was to assess the ability of the CellROX® Deep Red dye to detect oxidative stress in bacteria and to monitor membrane integrity using SYTOX® Blue. Specifically, FCM was used to assess the effect of various concentrations of hydrogen peroxide on Enterococcus faecalis and Fusobacterium nucleatum. The capacity of the combination of CellROX® Deep Red and SYTOX® Blue to distinguish non-oxidized (CellROX® Deep Red negative cells) from oxidized (CellROX® Deep Red positive cells) and membrane-damaged bacteria (SYTOX® Blue positive cells) was further investigated. Go to Material and Methods Bacteria A Gram-positive bacterium, Enterococcus faecalis (E. faecalis 135737, culture collection of the University Hospitals of Geneva, CH) and a Gram-negative bacterium, Fusobacterium nucleatum (Orale Mikrobiolgie Zurich culture collection - OMZ 598) have been used in this study. E. faecalis and F nucleatum were cultured from frozen stocks onto Columbia and Schaedler agar plates respectively. Bacteria from agar cultures were transferred in liquid media and incubated overnight at 37 °C (stationary phase). Brain heart infusion has been used as liquid media for E. faecalis and Schadler broth for F. nucleatum (all from Oxoid AG, Pratteln, Switzerland). Cultures of F. nucleatum were maintained under anaerobic conditions using packs of carbon dioxide (GasPack Anaerobe Gas Generating Pouch system with indicator, Becton Dickinson). On the day of the experiment bacteria were centrifuged and re-suspended in NaCl 0.9% to spectrophotometrically adjust the bacterial concentration at OD,600nm 0.2: ~1.2 x107 bacteria, Biowave II,’ Biochrom WPA, Cambridge, GB). Viability curve Hydrogen peroxide 3% w/w (Bichsel AG, Interlaken, Bern, Switzerland) has been used to induce oxidative stress in bacteria. To ensure that oxidative stress was measured on live cells, a H2O2 dose-response was determined for both strains using the LIVE/DEAD BacLight Bacterial Viability kit (Life technologies, Switzerland) and the Accuri C6 flow cytometer as previously described (BD Accuri Cytometers, Ann Arbor, USA) [16,17]. Briefly, 100μL of bacterial suspension have been mixed with 100μL of hydrogen peroxide at different concentrations (from 0 to 980mM in samples) and incubated at 37 °C for 15min before staining with the LIVE/DEAD solution. A sublethal range of H2O2 was defined as concentrations causing less than 10% cell death. Defining a labeling concentration of CellROX® Deep Red for bacteria To define an optimal labeling concentration of CellROX® Deep Red (Life technologies, Switzerland) for E. faecalis and F.nucleatum, bacteria have been exposed to maximal sublethal H2O2 concentrations (as defined by the viability curve), during 15min at 37 °C. Cultures were then incubated with various concentrations of CellROX® Deep Red (0μM, 0.5μM, 1μM, 2μM, 4μM and 8μM) during 30min. After incubation, bacteria were sonicated 20sec to disperse aggregates (35kHz, Sonoroex, Bandelin electronics, Berlin, DE) and fixed with 3.7% formaldehyde (VWR International AG, Dietikon, Switzerland). Fixation has been performed for biosafety reasons. Testing increasing H2O2concentrations To verify that the CellROX® Deep Red fluorescence increased accordingly with oxidative stress, bacterial samples were exposed to different concentrations of H2O2 (from 0mM to 160mM in samples) for 15min at 37 °C. The samples have then been incubated with 4μM CellROX® Deep Red (previously determined working concentration) during 30min at 37 °C, sonicated 20sec and formaldehyde fixed. Control samples were labelled with 4μM CellROX® Deep Red without prior exposure to H2O2. Dual labeling with CellROX® Deep Red and SYTOX® Blue To test whether the combination of CellROX® Deep Red and SYTOX® Blue would identify different bacterial oxidative states, samples of E. faecalis and F. nucleatum have been exposed for 15min to different concentrations of H2O2 (0mM- 980mM in samples) and then incubated with 4μM CellROX® Deep Red for 30min at 37 °C. Controls including non-oxidized (0mM H2O2), oxidized (maximum sublethal H2O2 concentration) and membrane-damaged (980mM H2O2) cells were used for comparisons. Samples were then sonicated 20sec, and SYTOX® Blue added to a final concentration of 4μM. Incubation with the SYTOX® Blue was continued for 15min. Experiments using both CellROX® Deep Red and SYTOX® Blue have been made on live cells (no formaldehyde fixation). Flow cytometric analysis Samples have been analyzed using a Gallios flow cytometer (Beckman Coulter, California, USA). Prior to analysis, unlabeled bacteria were run to optimize voltage settings (trigger on the FSC, voltage 650 on the FL-6 channel, 500 on the FL-9), and flow rate was set on low. Bacterial events were discriminated from debris using forward (FSC-A) and side scatter (SSC-A). Doublets have been excluded for analysis by FSC-height and width. CellROX® Deep Red signal (excitation/emission; 644/665) was collected in the FL6 channel (BP 660/20) and the SYTOX® Blue signal (excitation/emission; 444/480) in the FL9 channel (BP 450/50). Gates applied for population discrimination were set manually based on control samples. Data were exported and analyzed with FlowJo software (FlowJo for Windows, version 10.0.06, 2014, Tree Star Inc., Ashland, Oregon, U.S.A.). The geometric mean of fluorescence intensities (MFI) was expressed as relative fluorescence units (RFU). Statistical analysis All experiments were performed in triplicate and repeated 3 times. Results were statistically analyzed using one-way analysis of variance (ANOVA) and Tukey multiple comparison intervals (α = 0.05). Go to Results Viability curve Concentrations of H2O2 up to 80mM and 40mM were respectively applied on E. faecalis and F. nucleatum without inducing more than 10% reduction in viability (data not shown). Therefore, these concentrations were defined as the maximum oxidative stress to be applied on E. faecalis and F nucleatum, and were retained to determine an optimum working concentration of CellROX® Deep Red for measuring oxidative stress. Defining a labeling concentration of CellROX® Deep Red for bacteria Control suspensions (0μM CellROX® Deep Red) of E. faecalis and F nucleatum respectively exposed to 80mM and 40mM H2O2 displayed a natural fluorescence around 300 RFU (Figure 1A & 1B). Concentrations of CellROX® Deep Red ranging from 0.5μM to 4μM resulted in increased fluorescence intensities in both bacterial species. At 4μM CellROX® Deep Red, fluorescence intensities reached 2807±945 RFU and 3104±1151 RFU for E. faecalis and F. nucleatum respectively. Further increasing CellROX® Deep Red concentrations to 8μM resulted in decreased fluorescence intensities, for both strains (Figure 1A & 1B). Therefore, 4μM has been selected as the optimal concentration of CellROX® Deep Red for measuring oxidative stress in E. faecalis and F. nucleatum. Testing increasing H2O2 concentrations Control cultures (0mM H2O2) labeled with 4μM CellROX® Deep Red displayed a fluorescence of 648±90 RFU for E. faecalis and 1138±396 RFU for F. nucleatum (Figure 1C & 1D). For both strains, increasing concentrations of H2O2 progressively increased CellROX® Deep Red signal. For E. faecalis, the fluorescence signal was maximal at 80mM H2O2 (3001±633 RFU), whereas F nucleatum exhibited maximal fluorescence intensities at 40mM H2O2 (3753±1575 RFU) (Figure 1C & 1D). These H2O2 concentrations corresponded to the maximum sublethal values of each strain. Increasing H2O2 concentrations above these values resulted in decreased CellROX® Deep Red fluorescence in both strains. Dual labeling with CellROX® Deep Red and SYTOX® Blue Flow cytometric analysis of non-oxidized controls (0mM H2O2) showed 96.2% of E. faecalis and 85.6% of F. nucleatum with low signals for both CellROX® Deep Red and SYTOX® Blue (Figure 2A & 2E). Oxidized controls (samples under maximal H2O2 stress) exhibited increased CellROX® Deep Red signal (Figure 2B & 2F). At 160mM H2O2, bacteria of both strains displayed increased SYTOX® Blue signal (Figure 2C & 2G). When exposed to 980mM H2O2, 97.1% of E. faecalis and 98.8% of F. nucleatum exhibited a SYTOX® Blue positive signal in addition to the CellROX® Deep Red fluorescence (Figure 2D & 2H). Notably, CellROX® Deep Red signal intensity decreased in E. faecalis at H2O2 concentrations above 80 mM (Figure 2B-2D). Go to Discussion In the current study, we successfully applied both the CellROX® Deep Red and the SYTOX® Blue to a Gram-positive and -negative bacterium to measure oxidative stress and to discriminate non-oxidized from oxidized and damaged cells. Concentrations of hydrogen peroxide selected to induce oxidative stress were in the millimolar range (40mM-80mM). Previous studies have shown that E. faecalis exposed 15min to H2O2 retains viability up to 15-20mM [18,19]. The higher concentrations reported in the current study may be attributed to the different methods used for viability measurement. Whereas FCM in combination with the LIVE/DEAD staining has been used in this study to evaluate membrane integrity, previous studies measured viability by culture plating techniques [20]. There is evidence showing that bacteria proliferation capacity is affected at lower concentrations of H2O2 than membrane integrity [21-23]. An optimal concentration of CellROX® Deep Red has been determined by testing several concentrations of dye on stressed bacteria. Results indicate that fluorescence increased when increasing concentrations of dye up to 4μM, but tended to decrease at 8μM (Figure 1A & 1B). This pattern of signal intensity suggests equilibrium between the dye and oxidants. At concentrations below 4μM, the dye would be saturated by oxidants and additional oxidative stress would not be detected by lack of free dye. Therefore CellROX® Deep Red at 4μM has been selected as the optimal labeling concentration in these bacterial strains. The tendency of fluorescence to decrease at concentrations above 4μM, may be explained by a phenomenon of FRET quenching since the excitation/emission spectra of the CellROX® Deep Red overlap (644/665nm) [24]. Samples labeled with 4μM CellROX® Deep Red and exposed to increasing oxidative stress displayed gradually increasing fluorescence signals with highest intensities at the maximal sublethal H2O2 concentrations. These data confirm that CellROX® Deep Red fluorescence increased accordingly with the oxidative stress applied. Notably, F nucleatum exhibited higher fluorescence intensities than E. faecalis at all H2O2 concentrations tested (Figure 1C & 1D). A possible explanation might be related to a higher uptake capacity of CellROX® Deep Red by F. nucleatum, since the bacterium has a larger cytoplasm; E. faecalis measures about 1μm while F. nucleatum reaches 5-25μm [25,26]. Also strict anaerobes as F. nucleatum, are more sensitive to oxidative stress probably due to enzymes particularly prone to oxidation, as those containing exposed glycyl radicals and low-potential iron sulfur clusters [27,28]. Reaction with these targets may lead to further oxidative reactions potentially responsible for higher ROS production and fluorescence signal in F. nucleatum. The combination of CellROX® Deep Red and SYTOX® Blue showed that control cells (0mM H2O2) displayed a double negative signal and were located in quadrant 4 (Figure 2A & 2E). On the contrary, oxidized controls (40mM and 80mM) displayed an increased CellROX® Deep Red signal, testimony of oxidative stress; these bacteria were located in quadrant 3 (Figure 2B & 2F). For both strains an uptake of SYTOX® Blue at H2O2 concentrations above sub lethal values (160mM) was observed, thereby confirming that membrane injury occurred under such elevated oxidative stress (Figure 2C & 2G). In E. faecalis, sub-populations characterized by different intracellular concentrations of dyes were observed, possibly indicating intermediate states of membrane injury (Figure 2C). Membrane- damaged controls (980mM) showed bacteria positive for both CellROX® Deep Red and SYTOX® Blue (Figure 2D & 2H). This is not surprising as the membrane-damage of these cells was produced by an excess of oxidative stress. However, E. faecalis cells lost CellROX® Deep Red fluorescence possibly due to a leakage of the dye from the cytoplasm, since membrane injury was shown to occur at such elevated H2O2 concentrations (Figure 2B-2D). The presence of an outer membrane in the Gram-negative bacterium, F. nucleatum, may have accounted for a better cytoplasmic retention of the CellROX® Deep Red dye. Gram-positive and -negative bateria have previously been shown to react differently to the same staining protocol [29]. Go to Conclusion The results of the current study indicate that the CellROX® Deep Red dye can be used to measure oxidative stress in E. faecalis and F. nucleatum. The combinational use of CellROX® Deep Red and SYTOX® Blue allowed the identification of nonoxidized, oxidized and damaged bacteria. Future studies seem warranted to assess the reactivity of the CellROX® Deep Red dye in presence of other oxidative challenges. Go to Acknowledgments This study was supported by Grant #31003A-149962 of the Swiss National Science foundation. Mr. Aubry- Lachainaye JP and Mrs. Gamerio C are acknowledged for their contribution during flow cytrometry analysis. The authors deny any conflicts of interest related to this study. To know more about Journal of Biotechnology and Microbiology Click here: https://juniperpublishers.com/aibm/index.php To know more about Juniper Publishers Click here: https://juniperpublishers.com/index.php
"CD" in immunology stands for Cluster of Differentiation and includes cell surface markers that can be detected by lab technique called flow cytometry. Well, we will just focus on the ones you need to remember
This blog post will take you through the various gating strategies for effective flow cytometry analysis.
Previous studies have suggested that carnivory and holokinetic chromosomes might be associated with genome downsizing in plants. Veleba et al. analysed
Data analysis in flow cytometry relies on the principle of gating. Here we show how gates and regions drawn on dot plots and histograms allow investigation and analysis of specific populations.
The main difference between flow cytometry and FACS is that flow cytometry allows to rapidly, accurately, and simply collect data related to many parameters
Chronic lymphocytic leukemia (CLL) is a blood cancer characterized by abnormal lymphocyte accumulation. Causes are unknown, and symptoms include fatigue and swollen lymph nodes. CLL diagnosis involves blood tests and biopsies. Treatments vary based on disease stage, with outcomes ranging from observation to therapy. 6 Must Tests for CLL Diagnosis Accurate CLL diagnosis is crucial in Chronic Lymphocytic Leukemia (CLL) to determine the stage, guide treatment decisions, and monitor disease progression, improving patient outcomes and quality of life. In the chronic lymphocytic leukemia diagnosis, various tests may be performed to evaluate the condition. Here is a list of common tests used: Complete Blood Count (CBC) Flow Cytometry Immunophenotyping Bone Marrow Aspiration and Biopsy Cytogenetic Analysis Fluorescence In Situ Hybridization (FISH) 1. Complete Blood Count (CBC) Comprehensive analysis of blood components for CLL diagnosis. Accurate evaluation of red and white blood cell counts. Assessment of hemoglobin levels for oxygen-carrying capacity. Category Details Also Known As Hemogram, Full Blood Count (FBC) Purpose Assess overall health, detect blood disorders Sample Blood sample Preparation None Procedure Blood draw from a vein Test Timing 2-4 hours Test Price (INR) 300-800 Result Value Quantitative measurement of blood components Normal Value Varies based on age, gender, and other factors Accuracy High Interpretation Evaluated by healthcare professionals based on specific needs In CLL diagnosis, Complete Blood Count (CBC) test assesses overall health, detects disorders/infections. Quick, accurate, and widely available at an affordable price. 2. Flow Cytometry Precise cellular analysis using powerful technology. Quantification of cell populations with high accuracy. Identification of specific cell types for CLL diagnosis purposes. Category Details Also Known As Immunophenotyping Purpose Analyze cell populations Sample Blood, bone marrow, or bodily fluids Preparation Depends on the specific sample type Procedure Cell staining & analysis using a flow cytometer Test Timing Same day results Test Price (INR) 2,000-6,000 Result Value Identification of cell types Normal Value Varies based on specific cell populations Accuracy High Interpretation Assessed by medical professionals based on context Flow Cytometry (Immunophenotyping) accurately analyzes cell populations for various samples, providing valuable CLL diagnosis insights. Quick results available (INR 2,000-10,000) with high accuracy and interpretation by medical professionals. 3. Immunophenotyping Accurate identification and characterization of specific immune cell types. Evaluation of cell surface markers to understand immune system function. Precise detection and classification of abnormal cells for chronic lymphocytic leukemia diagnosis and monitoring. Category Details Also Known As Flow Cytometry Purpose Identify immune cell types Sample Blood, bone marrow, bodily tissues Preparation Depends on the sample type Procedure Cell staining & analysis using a flow cytometer Test Timing Usually same day results Test Price (INR) 2,000-10,000 Result Value Characterization of immune cells Normal Value Varies on specific immune cell populations Accuracy High Interpretation Assessed by medical professionals based on context Immunophenotyping (Flow Cytometry) identifies and characterizes immune cell types in samples like blood or tissue. Quick results available (INR 2,000-10,000) with high accuracy, aiding in CLL diagnosis and medical interpretation. 4. Bone Marrow Aspiration and Biopsy Diagnostic procedure for evaluating bone marrow health and detecting abnormalities. Collection of bone marrow samples for microscopic examination and genetic analysis. Accurate CLL diagnosis of hematopoietic cells, identifying blood disorders and malignancies. Category Details Also Known As Bone Marrow Examination Purpose Assess bone marrow health & blood disorders Sample Bone marrow from hip bone Preparation None Procedure Extracting liquid marrow Test Timing A few days Test Price (INR) 3,000-8,000 Result Value Microscopic examination Normal Value Varies based on age, gender Accuracy High Interpretation Assessed by healthcare professionals based on specific needs Bone Marrow Aspiration and Biopsy test evaluates bone marrow health, detects abnormalities, and blood disorders. Quick and accurate chronic lymphocytic leukemia diagnosis results (INR 3,000-8,000) aid medical interpretation by professionals. 5. Cytogenetic Analysis Detailed examination of chromosomes to detect genetic abnormalities and chromosomal rearrangements. Identification of specific genetic mutations and alterations related to various diseases. Precise CLL diagnosis for personalized treatment planning and prognosis assessment. Category Details Also Known As Chromosome Analysis Purpose Detect genetic abnormalities, chromosomal rearrangements Sample Blood, bone marrow, or tissue samples Preparation Depends on the sample type Procedure chromosomes microscopy & genetic techniques Test Timing A few days Test Price (INR) 5,000-15,000 Result Value Identification of genetic mutations, chromosomal alterations Normal Value Varies based on genetic markers Accuracy High Interpretation Assessed by medical professionals based on specific needs In CLL diagnosis, Cytogenetic Analysis detects genetic abnormalities and rearrangements. Results (INR 5,000-15,000) aid interpretation of genetic mutations and chromosomal alterations by medical professionals. 6. Fluorescence In Situ Hybridization (FISH) Utilizes fluorescent probes to detect specific DNA sequences and chromosomal abnormalities. High sensitivity and specificity for identifying genetic alterations associated with diseases. Aids in chronic lymphocytic leukemia diagnosis, prognosis, and treatment decisions for various genetic disorders and cancers. Category Details Also Known As FISH Purpose Detect specific DNA sequences Sample Cell Sampling Preparation Depends on the sample type Procedure DNA probe binding to specific sequences Test Timing A few days Test Price (INR) 5,000-15,000 Result Value Identification of specific DNA sequences Normal Value Varies on specific genetic markers Accuracy High Interpretation Assessed by medical professionals based on specific needs Fluorescence In Situ Hybridization (FISH) test detects specific DNA sequences and chromosomal abnormalities. Results (INR 5,000-15,000) aid interpretation by identifying genetic markers accurately and reliably. Chronic Lymphocytic Leukemia Tests Overview Test Name Complete Blood Count (CBC) Flow Cytometry Immunophenotyping Also Known As Hemogram, Full Blood Count Immunophenotyping Flow Cytometry Purpose Detect blood disorders & infections Identify different cell populations Determine cell markers for CLL diagnosis Sample Preparation Blood sample in EDTA tube Blood or bone marrow sample Blood or bone marrow sample Procedure Automated blood analyzer Flow cytometry analysis Flow cytometry analysis Test Timing 2-4 hours Same day results Same day results Test Price (INR) 300-800 2,000-10000 2,000-6,000 Result Value Quantitative measurement of blood components Identification of cell types Characterization of immune cells Normal Value Varies by age Varies Varies Accuracy High High High Interpretation Evaluated by healthcare professionals based on specific needs Assessed by medical professionals based on context Assessed by medical professionals based on context *Test Price, range, and timing may vary as per location, lab type, and procedure. Complete Blood Count (CBC), Flow Cytometry, and Immunophenotyping are vital CLL diagnosis tests used to assess health, detect disorders, and determine cell markers. They involve collecting samples and utilizing analysis techniques, with varying result values, prices, and interpretations. Chronic Lymphocytic Leukemia Differential Diagnosis Similar Diseases Differentiating Factors Small Lymphocytic Lymphoma Bone marrow biopsy and lymph node involvement Hairy Cell Leukemia Presence of hairy cells and associated markers Mantle Cell Lymphoma Cyclin D1 expression and characteristic t(11;14) translocation Prolymphocytic Leukemia Lymphocyte morphology, CD5 expression Acute Lymphoblastic Leukemia Blasts in peripheral blood and bone marrow Lymphoplasmacytic Lymphoma Presence of monoclonal IgM and MYD88 mutation Follicular Lymphoma Lymph node architecture and t(14;18) translocation Marginal Zone Lymphoma Lymph node architecture and specific immunophenotypic markers T-Cell Prolymphocytic Leukemia T-cell phenotype, lymphocyte morphology Splenic Marginal Zone Lymphoma Lymph node architecture, splenic involvement Chronic Lymphocytic Leukemia can be distinguished from similar diseases through factors such as lymph node involvement, presence of specific markers, cytogenetic abnormalities, and morphology. Differential diagnosis includes Small Lymphocytic Lymphoma, Hairy Cell Leukemia, Mantle Cell Lymphoma, and others. Best Doctor for Chronic Lymphocytic Leukemia Specialist Description Hematologist-Oncologist Specializes in CLL and blood cancers Medical Oncologist Expert in cancer treatment Hematopathologist Focuses on blood disorders For CLL, The best specialist is a Hematologist-Oncologist, who specializes in CLL, including blood cancers. 7 Interesting Facts of Chronic Lymphocytic Leukemia (CLL) Diagnosis CLL is often detected incidentally in routine blood tests. It is more common in older individuals. Some patients with CLL may remain asymptomatic for years. Genetic abnormalities play a role in CLL diagnosis and development. CLL is not always considered a cancer that requires immediate treatment. CLL can affect lymph nodes, bone marrow, and other organs. CLL patients may experience periods of remission and relapse. Conclusion Accurate chronic lymphocytic leukemia diagnosis is crucial in Chronic Lymphocytic Leukemia (CLL). Tests like CBC, flow cytometry, and more help determine treatment and predict outcomes. Early detection and comprehensive evaluation lead to better CLL diagnosis. Reference Chronic lymphocytic leukemia - Wikipedia [1]. Chronic Lymphocytic Leukemia - StatPearls - NCBI Bookshelf [2].
Definition, Principle, Instrumentation/Parts, Protocol/Procedure/Process/Steps, Types, Applications/Uses, Limitations of Flow Cytometry.
Fluorescence levels and light scatter as particles pass through the lasers provide information about the particle’s properties. Filters control specificity.
Data analysis in flow cytometry relies on the principle of gating. Here we show how gates and regions drawn on dot plots and histograms allow investigation and analysis of specific populations.
T- cells are the mediators of cell mediated immune response. T cells originate in bone marrow and mature in thymus. T cells interaction wi...
View our interactive Hematopoietic Stem Cell Differentiation Pathways Lineage-specific Markers.
Gates and regions can be added to flow cytometry dot plots and histograms to identify specific populations based on FSc, SSc and fluorescence. Find out more
The acquired immune response or adaptive immune response operates through two types of cells specifically B-cells and T-cells. 4 cell s...
Journey through the body's unsung heroes with our guide on the lymphatic system's anatomy and physiology. Nursing students, delve deep into the silent networks that defend and detoxify our body every moment.
76 cm x 61 cm 30 " x 24" 2012 Urethane and acrylic binders, pigments in dispersal water, dry iridescent pigments and resin on panel. Sold
Acute Lymphoblastic Leukemia (ALL) is a malignant proliferation of lymphoblasts in the blood and bone marrow.
Did your heart just skip a beat? Then you must love this heart anatomy print. This Valentines day print is a great gift for any one that you love who loves human anatomy! ART WORK STATS: • paper size: available in two sizes: 8 in X 10 in or 11 in x 14 in • printed on Aurora Fine Art Natural (100% cotton rag, acid-free, matte) • printed with Epson archival pigment inks • copy right watermark removed before printing SHIPPING INFO: • print will be packed in a sealed clear cello bag with a thick backing board, and shipped inside a rigid protective envelope • proof of mailing with each item is provided © 2013 Rachel Ignotofsky All rights reserved. Usage, reproduction, or altering of artwork is not permitted without permission. Each piece of artwork is an intellectual property and is protected by the copyright law.
