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Cytodiagnosis

Cytodiagnosis: The practice of diagnosing disease through the microscopic examination and analysis of cells.
This discipline leverages cellular morphology, biochemical and molecular characteristics to identify pathological conditions.
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Most cited protocols related to «Cytodiagnosis»

Improving Systematic Review Efficiency Through Deduplication

Cited 18 times

The Systematic Review Assistant-Deduplication Module (SRA-DM) project was developed in 2013 at the Bond University Centre for Research in Evidence-Based Practice (CREBP). The project aimed to reduce the amount of time taken to produce systematic reviews by maximising the efficiency of the various review stages such as optimising search strategies and screening, finding full text articles and removing duplicate citations.
The deduplication algorithm was developed using a heuristic-based approach with the aim of increasing the retrieval of duplicate records and minimising unique records being erroneously designated as duplicates. The algorithm was developed iteratively with each version tested against a benchmark dataset of 1,988 citations. Modifications were made to the algorithm to overcome errors in duplicate detection (Table 1). For example, errors often occurred due to variations in author names (e.g. first-name/surname sequence, use/absence of initialisation, missing author names and typographical errors), page numbers (e.g. full/truncated, or missing), text accent marks (e.g. French/German/Spanish) and journal names (e.g. abbreviated/complete, and ‘the’ used intermittently). The performance of the SRA-DM algorithm was compared with EndNote’s default one step auto-deduplication process. To determine the reliability of SRA-DM, we conducted a series of validation tests with results of different literature searches (cytology screening tests, stroke and haematology) which were retrieved from searching multiple biomedical databases (Table 2).Table 1

SRA-DM algorithm changes

Iterations	Changes to algorithms
First iteration	Matching criteria were based on simple field comparison (ignoring punctuation) with checks against the year field since this field has a lower probability for errors because it is restricted to integers 0–9 and therefore the best non-mistakable field.
Second iteration	Short format page numbers were converted to full format (e.g. 221–226, 221–6), and the algorithm was further modified to increase the sensitivity by incorporating matching criteria on authors OR title.
Third iteration	Match author AND title with the extension of the non-reference fields from only ‘year’ to year OR volume OR edition.
Fourth iteration	The fourth algorithm extended the matching criteria of the third algorithm, with the addition of an improved name matching system. This was context aware of author name variations, i.e. initialisation, punctuation and rearranged author listings using fuzzy logic, so that differences could be accommodated. For example, the following names are all syntactically equivalent and will match as identical authors:
	1. William Shakespeare
	2. W. Shakespeare
	3. W Shakespeare
	4. William John Shakespeare
	5. William J. Shakespeare
	6. W. J. Shakespeare
	7. W J Shakespeare
	8. Shakespeare, William
	9. Shakespeare, W
	10. Shakespeare, W, A
	11. Shakespeare, W, A, B, C
	12. William Shakespeare 1st
	13. William Shakespeare 2nd
	14. William Shakespeare IV
	15. William Adam Bob Charles Shakespeare XVI

Table 2

Databases searched for retrieval of citations for validation testing

Datasets	Databases searched
Cytology screening tests dataset	1. Cochrane Controlled Trials Register (CCTR)
	2. Cochrane Database of Systematic Reviews (CDSR)
	3. EMBASE
	4. MEDLINE
	5. National Research Register (NRR)
	6. Database of Assessments of Reviews of Effectiveness (DARE)
	7. NHS Health Technology Assessment (HTA)
	8. PreMEDLINE
	9. Science Citation Index
	10. Social Sciences Citation Index
Haematology dataset	1. MEDLINE
	2. EMBASE
	3. MEDLINE In-Process
	4. Biological Abstracts
	5. NHS Health Technology Assessment (HTA)
	6. Cochrane Controlled Trials Register (CCTR)
	7. Cochrane Database of Systematic Reviews (CDSR)
	8. CINAHL
	9. Science Citation Index
	10. Social Sciences Citation Index
Stroke dataset	1. MEDLINE
	2. EMBASE
	3. CENTRAL
	4. CINAHL
	5. PsycInfo

Rathbone J., Carter M., Hoffmann T, & Glasziou P. (2015). Better duplicate detection for systematic reviewers: evaluation of Systematic Review Assistant-Deduplication Module. Systematic Reviews, 4(1), 6.

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Publication 2015

Accent Biopharmaceuticals Cerebrovascular Accident Cytodiagnosis Hispanic or Latino Hypersensitivity Technology Assessment, Biomedical

Cervical Lesions Screening Cohorts in China

Cited 6 times

All clinical samples in this study were organized using the Fujian Province Cervical Lesions Screening Cohorts, Fujian, P.R. China. In one of these cohorts, 19,446 women assessed using Cervista^® HR-HPV assay and the Thinprep^® Cytologic Test at the Fujian Provincial Maternity and Children’s Hospital between January 2012 and December 2016 were enrolled. The population eligible for this study involved two arms, one consisting of healthy patients undergoing routine physical examination and another consisting of patients visiting the outpatient clinic for any gynecologic conditions other than cancer. Qualified participants were those for whom HPV was positively detected by the Cervista^® assay, with definite cervical histopathology results, and who had a complete follow-up record (Figure 1). In total, 698 eligible women who were histopathologically abnormal were classified in the case group. Another 698 women who were age matched with the case group and who were histopathologically normal were placed in the control group. All participants were examined, followed, and diagnosed using standardized protocols. All participants in this study provided written informed consent. The protocol was reviewed and approved by the Ethical Committees of the Fujian Provincial Maternity and Children’s Hospital.

Dong B., Sun P., Ruan G., Huang W., Mao X., Kang Y., Pan D, & Lin F. (2018). Type-specific high-risk human papillomavirus viral load as a viable triage indicator for high-grade squamous intraepithelial lesion: a nested case– control study. Cancer Management and Research, 10, 4839-4851.

Publication 2018

Age Groups Arm, Upper Biological Assay Cytodiagnosis Female Genital Diseases Malignant Neoplasms Neck Patients Physical Examination Woman

Functionalized SERS Nanoparticle Conjugation

Cited 6 times

The NPs (Cabot Security Materials Inc, Mountain View, CA) utilize a gold core and are encapsulated in silica. The gold-core diameters are ~60 nm and the overall dimension is ~120 nm. Additional details may be found in the literature^{9 (link)}. The silica surface of the NPs are functionalized with thiols to allow conjugation to a variety of targeting molecules (Fig. 6a). Three “flavors” of NPs were used, identified as S420, S421 and S440, each of which emits a characteristic Raman spectrum (Fig. 5b).
Stock concentrations of SERS NPs (800 pM in water) were diluted in 10 mM MOPS (Sigma-Aldrich, part No. M1254) buffer, pH 7.25, at a volume ratio of 1:1 (e.g. 200 μL buffer to 200 μL NPs in water), and then reacted with Cyto 647-maleimide (Cytodiagnostics Inc, part No. NF647-3-01), at 4.5 × 10⁵ molar equivalents per NP, at room temperature for 1 h to conjugate Cyto 647 fluorophores to the NP surface (Fig. 6a). Three different monoclonal antibodies (mAb) were used for conjugation: anti-EGFR “panitumumab” (Vectibix, NDC 55513-954-01), anti-HER2 (Thermo Scientific, MS-229-PABX), or an isotype control mAb (Thermo Scientific, MA110407). These mAbs were first eluted over a desalting spin column (Thermo Scientific, 89882) and buffer-exchanged into the MOPS buffer to remove preservatives such as sodium azide. All mAbs were purchased free of protein stabilizers such as BSA or gelatin. Antibodies were added to fluorescent NPs at 500 molar equivalents per NP, along with the heterobifunctional PEG cross linker SM(PEG)^{12 (link)} (Thermo Scientific, 22112), at 1.5 × 10⁴ molar equivalents per NP, and incubated at room temperature for 3 h. Following the primary conjugation reaction, MM(PEG)^{12 (link)} (Thermo Scientific, 22711), at 6 × 10⁵ molar equivalents per NP, was then added to the NPs and reacted over night at 4°C to block residual thiols on the NPs. Note that all reagents were degassed with dry argon gas prior to use and all reactions were conducted under anhydrous conditions in light-protected amber vials (Fisher Scientific, 03-391-36&03-391-17) on a vortex mixer (Fisher Scientific^™ Microplate Vortex Mixers, 02-216-101) set at 800 rpm. Finally, the NPs were reacted at room temperature with 2-mercaptoethanesulfonate (Sigma-Aldrich, M1511) for 30 min, at 9 × 10⁵ molar equivalents per NP, and the reaction mixture was then diluted at a volume ratio of 1:1 with storage buffer (20 mM MOPS at pH 7.5 with 0.1% BSA (Jackson Immuno, 001-000-162) and 0.05% sodium azide (Sigma-Aldrich, S2002)). The diluted reaction mixture was purified four times via centrifugation (1000 g for 10 min), in which the supernatant was removed and replaced with storage buffer after each round of centrifugation. The conjugated NPs were stored at 4°C and protected from light. UV-VIS spectrophotometry was used to measure the final concentration of the conjugated NPs (Supplementary Fig. 2).

Wang Y.“., Khan A., Som M., Wang D., Chen Y., Leigh S.Y., Meza D., McVeigh P.Z., Wilson B.C, & Liu J.T. (2014). Rapid ratiometric biomarker detection with topically applied SERS nanoparticles. Technology, 2(2), 118-132.

Publication 2014

2-Mercaptoethanesulfonate Amber Antibodies Argon Buffers Cardiac Arrest Centrifugation Cytodiagnosis EGFR protein, human Enzyme Multiplied Immunoassay Technique ERBB2 protein, human Flavor Enhancers Gelatins Gold Immunoglobulin Isotypes Light maleimide Molar Monoclonal Antibodies morpholinopropane sulfonic acid NP 647 Panitumumab Pharmaceutical Preservatives Proteins SAMHD1 protein, human Secure resin cement Silicon Dioxide Sodium Azide Spectrophotometry Sulfhydryl Compounds Vectibix

Impact of Jade Goody's Illness on Cervical Screening Attendance

Cited 6 times

The Exeter database holds information about all invitations and cytology tests on women invited for cervical screening. In England, women aged between 25 and 49 are invited every three years and those aged between 50 and 64 every five years. This is a recent change from screening women aged 20–64 ‘at least five-yearly’, which was variously interpreted locally as every three, four or five years. Annual and monthly data about attendances were obtained from 2004 onwards. The information extracted included women's age, region of residence, category of invitation, screening status (i.e. whether they had been previously invited or screened and, if so, how recently) and test result.
The data were extracted using the definitions for creating the tables in the annual statistical bulletin, with one exception. Those women who have screening tests without being prompted by the programme are counted in the bulletins as having been tested outside the screening programme. For this analysis, all women are counted as having been tested within the programme, other than those invited privately or those with no prior recall type (i.e. those not yet called).
The extracts on which this analysis is based were taken in October/November 2010. As the databases are continually being updated the numbers of women whose data were extracted differs slightly from corresponding numbers in the published bulletins; in all years, the numbers in these extracts are the higher – in 2008–09 by about 40,000 attendances, or 1% of the total.
The number of attendances routinely varies from month to month. In order to estimate the excess of tests taken in the months affected by the publicity surrounding Jade Goody's diagnosis and death, Poisson regression models were fitted to the counts for all other months assuming that the numbers of attendances are influenced both by a seasonal effect and a smoothly varying time effect. Different models were fitted to different age-groups and different categories of test (e.g. type of invitation, time since last test, etc.). The seasonal effect was modelled by allowing a separate multiplicative factor for each calendar month and the time effect was modelled in Stata as a natural cubic spline.⁵ Within the model, terms for other factors (e.g. age and invitation type) were also included. The differences between the observed numbers in the months of particular interest and those predicted by the model (applied to all other months) were taken to be the excess in those months. These residuals could either be positive or negative, but on average they are zero for the months on which the models are fitted. It is their values in the months surrounding Jade Goody's diagnosis and death that are taken as the estimated extra number of attendances.
Analysis was performed to examine whether there were variations in the response to the publicity by age, reason for/category of test, time since last test (where expected date of testing varies according to reason for test) and region. In order to estimate the numbers of women tested who might never have returned to the screening programme without the publicity about a celebrity's illness and death, it was assumed that all tests for such women were included among the excess tests; that all the tests occurred at least six months beyond the programme standard for early repeat tests, and beyond 60 months for routine recall; and that the distributions by time since last test for excess tests of all other women were similar to the respective usual underlying distributions by time since last test.
Published data from colposcopy were also examined following anecdotal reports of women who had previously failed to attend for follow up of an abnormal cytology result now reinstating their appointments and attending. These data are not held on the Exeter database. Finally, we analysed press coverage of the events in order to attempt to correlate screening activity with the publicity.

Lancucki L., Sasieni P., Patnick J., Day T, & Vessey M. (2012). The impact of Jade Goody's diagnosis and death on the NHS Cervical Screening Programme. Journal of Medical Screening, 19(2), 89-93.

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Publication 2012

A-factor (Streptomyces) Age Groups ARID1A protein, human Colposcopy Cuboid Bone Cytodiagnosis Cytological Techniques Diagnosis factor A Mental Recall Neck Woman

Cervical Cancer Screening Cotests Analysis

Cited 4 times

With the permission of Kaiser and National Cancer Institute Institutional Review Boards, results of all Pap smears with HPV DNA tests collected within 7 days of one another (“cotests”) between January, 2003 and April 2008 were assembled and tabulated from the laboratory databases. HPV DNA testing for 13 carcinogenic HPV genotypes (HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 68) was performed using the Food and Drug Administration-approved Hybrid Capture 2 (hc2; Qiagen Corporation, Gaithersburg, MD, USA) according to the manufacturer’s instructions. Cotests were excluded if the HPV testing was performed in duplicate or no result was available, or the Pap result was unsatisfactory or “other” (the 2001 Bethesda System general categorization used for “exfoliated endometrial cells present in a woman >40”). Paps that were interpreted as atypical squamous cells of undetermined significance (ASC-US), the threshold for referral to colposcopy (without an HPV test)(11 (link);17 (link)) or more severe cytology (≥ASC-US) were considered screen positives.
During the study period 812,598 cotests were identified in 580,289 women age 30 and older, of which 1,376 cotests were excluded for HPV results (none available or duplicate), and 13,295 cotests were excluded for Pap results of “other” or “unsatisfactory”, leaving 797,927 cotests (98.2%) for analysis. These data included multiple cotests from some women, the results of which were included in the analysis because the primary focus was the screening results in the general population over a period of time rather than the point prevalence.
Individual and paired test results were compared to patient age, categorized into 5-year age groups from age 30 to 79 and grouping all women 80 older into an 80 and older (80+) category. Cotesting was done according to KPNC cervical cancer screening guidelines (18 (link)), including the screening of a small percentage of women 65 and older (approximately 6.5% of all women cotested), for whom screening is recommended if they have either not had three or more documented, consecutive, technically satisfactory normal/negative cervical cytology tests after the age of 55 or had an abnormal/positive cytology test for which HPV testing was indicated. Continued screening is also left to the discretion of the clinician if a woman 65 or older tests HPV positive. These guidelines are consistent with national screening guidelines that recommend discontinuing cervical cancer screening above the age of 65 (http://www.ahrq.gov/) or 70 (18 (link)) only when these women have had adequate recent screening with normal Pap smears and are not otherwise at high risk for cervical cancer.
Prevalence ratios (HPV/Pap) with 95% confidence intervals (95%CI) were calculated to show the relative contribution of HPV to Pap for screening positive by cotesting. McNemar’s chi square test was used to test for statistical differences (p < 0.05) for testing HPV positive versus testing positive by Pap smears for all women and by age group. Binomial exact 95%CI were calculated for the overall and age group-specific HPV prevalence. NCSS 2004 (Kaysville, Utah) was used for statistical analyses.

Castle P.E., Fetterman B., (ASCP) S., Poitras N., Lorey T., Shaber R, & Kinney W. (2009). Five-Year Experience of Human Papillomavirus DNA and Papanicolaou Test Cotesting. Obstetrics and gynecology, 113(3), 595-600.

Publication 2009

Age Groups Atypical Squamous Cells of Undetermined Significance Cancer Screening Cells Cervical Cancer Colposcopy Cytodiagnosis Cytological Techniques Endometrium Ethics Committees, Research Genotype Human papillomavirus 16 Human Papillomavirus DNA Tests Hybrids Neck Patients Woman Zinostatin

Most recents protocols related to «Cytodiagnosis»

Conjugating Pathogenic Fibrils to Nanogold Beads

Timing: 1 day

In this section, we explain the protocol used to conjugate PFFs to 5 nm nanogold beads (Cytodiagnostics). We altered and optimized the standard conjugation protocol provided by the manufacturer (see product sheet on the manufacturer’s website: https://www.cytodiagnostics.com/collections/NHS-Activated-Gold-Nanoparticles).

Note: Since PFFs are very susceptible to aggregation, the nanogold-PFFs need to be administered to cells immediately after conjugation. Prepare the cells such that they are ready to coincide with the conjugation of PFFs to nanogold. Following the administration of PFFs to cells, proceed to prepare grids to characterize the nanogold-PFFs.

Note: all of step 1 should be performed in a tissue culture hood.

The conjugation of PFFs with 5 nm nanogold.a.

Add 48 μL of 5 mg/mL PFFs to 60 μL of the reaction buffer provided with the nanogold conjugation kit.i.

Thaw reaction buffer for 1 h on ice.

ii.

Retrieve 48 μL of 5 mg/mL PFFs (stored at −80°C) and immediately add 60 μL of ice-cold reaction buffer to the aliquot.

iii.

Incubate for 2 min at room temperature.

iv.

Slowly pipette the mixture up and down.

Add the reaction buffer-PFFs mixture to 1 vial of 5 nm nanogold.

Incubate for 1 h at 18°C.

Add 10 μL of quencher buffer (provided by the manufacturer) to the mixture.i.

Slowly pipette up and down using a p10 pipette set at 10 μL.

ii.

Avoid creating bubbles in the solution.

Incubate at room temperature for 5 min.

Note: the amount of reaction buffer and quencher buffer used, the amount and concentration of protein used, are all specified by the manufacturer (Cytodiagnostics). The steps above are an adaptation of their protocol for our specific conjugation assay.

Note: for PFFs visualization (labeled with nanogold beads, use a magnification between ×30,000 and ×68,000).

CRITICAL: Avoid rapid pipetting of the samples or sudden movements of the experimental microtube as they will aid in protein aggregation.

Characterization of conjugated PFFs.a.

Prepare a diluted sample of PFFs with a 1 mg/mL concentration.i.

Dilute using ddH₂O.

Place one drop of diluted PFFs on a carbon/formvar covered copper grid and allow the sample to sit for 10 min at room temperature.i.

Using a Whatman filter paper, remove the drop from the grid.

Fix the PFFs sample on the grid using 2% PFA (retrieved immediately from storage at 4°C).i.

Place one drop onto the grid and let the sample sit for 10 min at room temperature.

ii.

Remove PFA drop using filter paper.

Add one drop of ddH₂O onto the grid for 1 min at room temperature.i.

Remove with filter paper.

ii.

Repeat once more.

Stain the grid with Uranyless.i.

Centrifuge 1 mL of Uranlyess at 14,000 × g.

ii.

add one drop of the supernatant to the grid and wait for 5 min at room temperature.

Wash grid with ddH₂O as detailed in d.

Remove the drop using a Whatman filter paper.

Let the grid sit overnight on parafilm at room temperature.

CRITICAL: place grids in a box/chamber for their storage overnight to avoid the addition of contaminants (such as bacteria, dust, etc.) to the EM grid.

Visualize using EM to assess and measure the length of fibrils.a.

See Figure 2 for an example:Figure 2

Electron micrograph of nanogold conjugated PFFs

Uranyless was used for staining. Arrowheads point to a few examples of conjugated nanogold beads. Arrows indicate optimal examples of nanogold-labeled PFFs: small size and the appropriate amount of conjugation (>1 and <20 beads per PFFs). Scale bar = 100 nm.

Criteria for successful/ideal PFFs conjugation:

The majority of PFFs should have a length of less than 100 nm.

Most PFFs should be conjugated with at least one nanogold bead.

Only 1 in 20 nanogold beads should be unconjugated (free-floating).

Bayati A., Luo W., Del Cid-Pellitero E., Fon E.A., Durcan T.M, & McPherson P.S. (2023). Visualization of α-synuclein trafficking via nanogold labeling and electron microscopy. STAR Protocols, 4(1), 102113.

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Publication 2023

Acclimatization Bacteria Biological Assay Buffers Carbon Cells Cold Temperature Copper Cytodiagnosis Formvar Gold MLL protein, human Movement Protein Aggregates Proteins Stains Strains Technique, Dilution Tissues

Older Cervical Cancer Treatment Outcomes

All patients with newly diagnosed, biopsy-proved cervical cancer treated at Peking Union Medical College Hospital from January 2000 to December 2020 were identified. The inclusion criteria were 1) aged ≥65 years; 2) squamous carcinoma, adenocarcinoma, and other histopathology like adenosquamous carcinoma; 3) stage IB-IIA (FIGO 2008 and FIGO 2018, according to primary diagnosis time); 4) treated by radical operation or radiotherapy; 5) with complete medical records. Patients were excluded if they 1) were accompanied by other malignancies like ovarian cancer and colorectal cancer or 2) had a follow-up time<3 months. The follow up lasted till December 2021. The study was approved by the institutional review board (S-K 2058).
The demographic characteristics, treatment data, clinicopathological parameters, and follow-up information were collected for each patient. The patients were divided into radiotherapy group (RT group) and operation group (OP group) according to their primary intervention. Radiotherapy included simple radiotherapy and concurrent chemoradiotherapy (CCRT, using cisplatin/paclitaxel); surgery treatment included radical hysterectomy (Querleu–Morrow classification, type C resection) and pelvic/para-aortic lymphadenectomy with/without adjuvant chemotherapy (paclitaxel+cisplatin or paclitaxel+carboplatin), with/without adjuvant radiotherapy. Demographic and clinicopathological characteristics included age, primary complaint, pathology, tumor grade, FIGO stage (all patients were updated to FIGO 2018 stage), tumor size, ThinPrep cytologic test (TCT) result, HPV status, comorbidities, treatment, and adverse effects (according to Common Terminology Criteria for Adverse Events Version 5.0). Follow-up information like the date of last follow-up, the date of recurrence, disease status, site of recurrence, the date of death, and cause of death were obtained via telephone calls and outpatient records. The primary outcome was overall survival (OS), and the secondary outcomes were progression-free survival (PFS) and adverse effects. PFS was calculated from the date of treatment to the date of progression, death, or the last follow-up. OS was calculated from the date of treatment to the date of death or the last follow-up. Recurrence was defined as a measurable lesion on imaging (computed tomography, magnetic resonance imaging, or positron emission tomography computed tomography) after the initial radical treatment.

Gu Y., Cheng H., Cang W., Chen L., Yang J, & Xiang Y. (2023). Comparison of oncological outcomes in elderly early-stage cervical cancer patients treated with radical surgery or radiotherapy: A real-world retrospective study with propensity score matching. Frontiers in Oncology, 13, 1019254.

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Publication 2023

Adenocarcinoma Adenosquamous Carcinoma Aorta Biopsy Carboplatin Cervical Cancer Chemotherapy, Adjuvant Colorectal Carcinoma Concurrent Chemoradiotherapy Cytodiagnosis Diagnosis Disease Progression Ethics Committees, Research Hysterectomy Lymph Node Excision Malignant Neoplasms Neoplasms Operative Surgical Procedures Outpatients Ovarian Cancer Paclitaxel Patients Pelvis Radiotherapy Radiotherapy, Adjuvant Recurrence Scan, CT PET Squamous Cell Carcinoma TP protocol X-Ray Computed Tomography

DNA-Origami Immobilization and Labeling

Flow chambers were assembled by sandwiching double-sided tape in between PEG-passivated glass slide and cover slip with a volume of around 10 µL. Flow chambers contained 1 mm holes at each end for buffer exchange. A tubing with syringe was inserted at one end and sealed with 5-min epoxy. A 20–200 µL pipette tip was inserted at the other end for introducing buffers. First, a 0.1 mg/mL solution of neutravidin (Invitrogen Inc.) in 3% BSA in PBS was introduced into the biotin-PEG/PEG passivated flow cell and incubated for 5 min. Excess neutravidin was washed off with PBS. 250 pM DNA-origami structures were added in 100 µl Buffer B + (5 mM Tris-HCl, 10 mM MgCl₂, 1 mM EDTA and 0.05% (vol/vol) Tween 20 at pH 8.0) per channel, incubated for 15 min and washed with 1 mL buffer C. Gold nanoparticles (Cytodiagnostics, cat: G-90-100) diluted 1:10 in buffer C were introduced and incubated for 5 min before washing with buffer C. The mIgG2a antibody, conjugated with the S1-handle with P3 docking site (S1-P3, Table S2) was incubated at a concentration of 60 nM in 0.5× buffer C + 1.5% BSA in PBS for 15 min. Excess mIgG2a-DNA conjugate was washed off with 2 × 200 µl 0.5x buffer C. For the negative control, instead of the mIgG2a, 60 nM of the S1-P3 handle alone was incubated for 15 min and the following steps remained the same. After washing off the primary probe, the κLC-Nb-7xR3 was added in 0.5x buffer C + 1.5% BSA and incubated for >45 min at RT. Excess κLC-Nb was removed by washing with 2 × 200 µl 0.5× buffer C, 10 min before imaging.

Ferapontov A., Omer M., Baudrexel I., Nielsen J.S., Dupont D.M., Juul-Madsen K., Steen P., Eklund A.S., Thiel S., Vorup-Jensen T., Jungmann R., Kjems J, & Degn S.E. (2023). Antigen footprint governs activation of the B cell receptor. Nature Communications, 14, 976.

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Publication 2023

Biotin Buffers Cells Cytodiagnosis Edetic Acid Epoxy Resins Gold Immunoglobulins Magnesium Chloride neutravidin Syringes Tromethamine Tween 20

Nanobody Labeling of Fixed B Cells

MACS-purified B1-8hi B cells were fixed with pre-heated (37 °C) 4% paraformaldehyde (PFA) for 30 min at RT. After fixation, cells were washed two times with PBS and transferred to 6-channel µ-Slide VI 0.5 glass bottom (Ibidi). Cells were immobilized on the glass surface by centrifugation at 1000 g for 15 min in a swinging bucket rotor using in-house built adaptors. Channels were rinsed with PBS to remove unbound cells and subsequently incubated with 0.1 M NH₄Cl₂ solution for 5 min followed by 2× wash steps. Cells were permeabilized with 0.1% Triton X-100 (Sigma-Aldrich) for 5 min at RT followed by 2x wash. To assure cells remained immobilized to the glass surface, the slide was centrifuged once at 1000 g for 10 min. Hereafter, cells were incubated with blocking buffer (PBS, 3% BSA, 1 mM EDTA, 0.02% Tween-20, and 0.05 mg/mL salmon sperm DNA) in the dark for 1 h at RT. The DNA-modified κLC-Nb was diluted to 30 nM in blocking buffer and added to the cells for incubation at 4 °C O/N. The following day, cells were washed 3x with PBS and centrifuged at 1000 g for 10 min prior to post-fixation with pre-heated (37 °C) 4% PFA for 5 min at RT and followed by 2x wash. Prior to imaging, gold nanoparticles (Cytodiagnostics, cat: G-90-100) diluted 1:3 in buffer C (PBS, 500 mM NaCl) were introduced and incubated for 5 min before washing with buffer C.

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Publication 2023

B-Lymphocytes Buffers Cells Centrifugation Cytodiagnosis Edetic Acid Gold paraform Salmo salar Sodium Chloride Sperm Triton X-100 Tween 20

Drosophila Hemi-Thorax Staining and Imaging

Drosophila hemi-thoraces were isolated and stained as described above with phalloidin Alexa488 (1:2000) and the two nanobodies coupled to either P1, P3, or PS3 oligos (about 50 nM) overnight. Before embedding the samples into the chamber, they were washed two times with PBS + 1% Triton. Hemi-thoraces were embedded as described above. Before assembling the chamber, the cover slip was treated with 90 nm diameter gold nanoparticles (cat#: G-90-100, Cytodiagnostics, 1:10 dilution into methanol). After assembling, the chamber was filled with imaging buffer containing the complementary P1, P3, or PS3 imaging oligos (see below for imaging conditions) and sealed with Picodent glue.

Schueder F., Mangeol P., Chan E.H., Rees R., Schünemann J., Jungmann R., Görlich D, & Schnorrer F. (2023). Nanobodies combined with DNA-PAINT super-resolution reveal a staggered titin nanoarchitecture in flight muscles. eLife, 12, e79344.

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Publication 2023

2',5'-oligoadenylate Buffers Chest Cytodiagnosis Drosophila Gold Methanol Phalloidine Technique, Dilution VHH Immunoglobulin Fragments

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Tween 20 is a non-ionic detergent commonly used in biochemical applications. It is a polyoxyethylene sorbitan monolaurate, a surfactant that can be used to solubilize and stabilize proteins and other biomolecules. Tween 20 is widely used in various laboratory techniques, such as Western blotting, ELISA, and immunoprecipitation, to prevent non-specific binding and improve the efficiency of these assays.

Hydroquinone by Merck Group

Sourced in United States, Germany, India, China, United Kingdom, Australia, Italy, Canada

Hydroquinone is a chemical compound used in various laboratory applications. It is a crystalline solid with the chemical formula C₆H₄(OH)₂. Hydroquinone is commonly used as a reducing agent, antioxidant, and in photographic development processes.

Silver acetate by Carolina Biological Supply

Silver acetate is a chemical compound with the formula CH3COOAg. It is a white crystalline solid that is used as a precursor for the synthesis of other silver compounds. Silver acetate has applications in various industries, including photography, electroplating, and the production of other silver-based products.

Acetic acid by Merck Group

Sourced in United States, Germany, United Kingdom, Italy, India, China, France, Spain, Switzerland, Poland, Sao Tome and Principe, Australia, Canada, Ireland, Czechia, Brazil, Sweden, Belgium, Japan, Hungary, Mexico, Malaysia, Macao, Portugal, Netherlands, Finland, Romania, Thailand, Argentina, Singapore, Egypt, Austria, New Zealand, Bangladesh

Acetic acid is a colorless, vinegar-like liquid chemical compound. It is a commonly used laboratory reagent with the molecular formula CH3COOH. Acetic acid serves as a solvent, a pH adjuster, and a reactant in various chemical processes.

Sodium hydroxide (naoh) by Merck Group

Sourced in Germany, United States, India, United Kingdom, Italy, China, Spain, France, Australia, Canada, Poland, Switzerland, Singapore, Belgium, Sao Tome and Principe, Ireland, Sweden, Brazil, Israel, Mexico, Macao, Chile, Japan, Hungary, Malaysia, Denmark, Portugal, Indonesia, Netherlands, Czechia, Finland, Austria, Romania, Pakistan, Cameroon, Egypt, Greece, Bulgaria, Norway, Colombia, New Zealand, Lithuania

Sodium hydroxide is a chemical compound with the formula NaOH. It is a white, odorless, crystalline solid that is highly soluble in water and is a strong base. It is commonly used in various laboratory applications as a reagent.

Thinprep by Hologic

Sourced in United States, New Zealand

The ThinPrep system is a laboratory equipment product that prepares patient samples for cytological examination. It is designed to process and prepare cellular samples in a standardized manner to improve the quality and consistency of specimen slides for analysis.

What are the different types or variations of Cytodiagnosis?

Cytodiagnosis encompasses several distinct techniques and approaches, including exfoliative cytology, fine needle aspiration (FNA), and liquid-based cytology. Exfoliative cytology involves the examination of cells that have naturally sloughed off from tissues, while FNA utilizes a small needle to extract cells from a specific area for analysis. Liquid-based cytology processes cellular samples in a liquid medium to enable more precise examination and preparation for downstream testing.

What are the common applications of Cytodiagnosis?

Cytodiagnosis is widely used in the detection and diagnosis of various cancers, including lung, thyroid, breast, and cervical cancer. It also plays a critical role in the evaluation of infectious diseases, inflammatory conditions, and endocrine disorders. By examining the cellular characteristics, clinicians can often identify the underlying pathological processes and guide appropriate treatment strategies.

What are some of the key challenges or limitations in using Cytodiagnosis?

One of the main challenges in Cytodiagnosis is the potential for sampling errors or inadequate cellularity, which can lead to false-negative results. Additionally, the interpretation of cytological findings requires specialized expertise and experience, as the morphological features can sometimes be subtle or overlapping between different conditions. Proper specimen preparation and handling are also critical to ensure the accuracy and reliability of the test results.

How can PubCompare.ai assist in the optimal use and comparison of Cytodiagnosis protocols?

PubCompare.ai's AI-driven platform can empower researchers to explore the latest Cytodiagnosis protocols from literature, preprints, and patents more efficiently. The platform's intuitive tools can help identify the most effective protocols based on key performance metrics, such as sensitivity, specificity, and reproducibility. By leveraging AI to pinpoint critical insights, PubCompare.ai can assist researchers in choosing the best Cytodiagnosis protocols that align with their specific research goals, ultimately enhancing the accuracy and reliability of their findings.

More about "Cytodiagnosis"

Cytodiagnosis, also known as cell-based diagnosis or cytopathology, is a critical field in medical diagnostics that involves the microscopic examination and analysis of cells to identify pathological conditions.
This discipline leverages cellular morphology, biochemical, and molecular characteristics to provide valuable insights into the underlying causes of various diseases.
Researchers and clinicians often utilize techniques like the ThinPrep cytologic test, which involves the preparation of thin, uniform cell samples for microscopic analysis.
Agents like Sodium phosphate dibasic, Bovine serum albumin, and Tween 20 may be used in the sample preparation process to optimize cellular preservation and staining.
The Vitrobot Mark IV is a commonly used automated plunge-freezing device that helps in the preparation of cryogenic samples for electron microscopy, a key tool in cytodiagnosis.
Other chemicals like Hydroquinone, Silver acetate, Acetic acid, and Sodium hydroxide may also be employed in various cytological staining and imaging protocols.
PubCompare.ai's AI-driven platform empowers researchers to explore the latest cytodiagnosis protocols from literature, preprints, and patents, allowing them to identify top products and optimize their research using intuitive tools.
By incorporating synonyms, related terms, and key subtopics, researchers can enhance their understanding of this important field and stay at the forefront of advancements in cell-based diagnostics.