Reticulocyte counting using flow cytometry.

نویسندگان

  • P R Nobes
  • A B Carter
چکیده

A flow cytometric method for the quantitation of reticulocytes was refined for routine laboratory use. Blood (2 microliters) is added to 2 ml of 0.4 microM thiazole orange in phosphate buffered saline, incubated at room temperature for 90 minutes, and analysed on a Coulter EPICS Profile flow cytometer, with gating for red cells on the basis of forward and right angled light scatter. Blood (2 microliters) is also incubated with phosphate buffered saline alone as an unstained control. The adult reference range (mean +/- 2 SD), established from 30 laboratory personnel, is 19.4-59.2 x 10(9)/l (0.2-1.6%). Comparison of this technique was made on 39 selected patient samples with visual counting of cells stained with brilliant cresyl blue. The correlation between the two methods was 0.99 with slope 0.96 and intercept 0.02. The precision of the automated technique in three subjects with reticulocyte counts of 0.12%, 1.84%, and 14.3% was 33.3%, 7.3%, and 1.4%, respectively (coefficient of variations). In three patients studied serially after intensive chemotherapy, in whom the reticulocyte count quantitated by routine visual methods approached zero (0-0.1%) for eight to 18 days, the automated counts varied between 0 and 0.5%. Flow cytometric reticulocyte counting is thus a simple and highly reliable methodology for the quantitation of normal and raised reticulocyte counts but cannot be reliably used to quantitate a subnormal level.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Optimized Method for Reticulocyte Counting: Simple, Accurate, and Comparable to Flow Cytometry

Background: Reticulocytes are immature red blood cells with RNA, spending the final stages of their maturation in the peripheral blood. The number of reticulocytes in the peripheral blood is the salient evidence of the effectiveness of bone marrow to produce red blood cells. Currently, reticulocyte count is done manually or automatically in clinical laboratories. Difficulties and limitations of...

متن کامل

Multivariate statistical modeling: alternative approach to test evaluation, applied to counting reticulocytes by flow cytometry.

We present a statistical path analysis model for the evaluation of two tests in the absence of a "gold standard" method. This model is applied to the evaluation of flow-cytometric and visual reticulocyte counting by using as the comparison method a combination of three hematological measurements: hemoglobin concentration (HGB), mean cellular volume (MCV), and erythrocyte density width (EDW). We...

متن کامل

Microsoft Word - NEF366BF

Dr. P. Zachée, Department of Hematology, University Hospital Leuven, Herestraat 49, B-3000 Leuven (Belgium) Table 1. Results of PVC, reticulocyte count, reticulocyte index and mean fluorescence before and after 3 months of rH-EPO treatment After rH-EPO p values Before rH-EPO Dear Sir, The enumeration of reticulocytes is the simplest, and perhaps the best way to assess effective red blood produc...

متن کامل

Biological evaluation of recombinant human erythropoietin in pharmaceutical products.

The potencies of mammalian cell-derived recombinant human erythropoietin pharmaceutical preparations, from a total of five manufacturers, were assessed by in vivo bioassay using standardized protocols. Eight-week-old normocythemic mice received a single subcutaneous injection followed by blood sampling 96 h later or multiple daily injections with blood sampling 24 h after the last injection. Re...

متن کامل

Development and Evaluation of a Novel In-Clinic Automated Hematology Analyzer, ProCyte Dx, for Canine Erythrocyte Indices, Leukogram, Platelet Counts and Reticulocyte Counts

A novel hematology analyzer for small animal medicine, ProCyte Dx, was developed from combination of the fluorescence laser flow cytometry and laminar flow impedance technologies, and its accuracy was evaluated by comparing with the conventional impedance-based hematology analyzer, pocH-100iV Diff, or microscopic manual cell counting methods with staining blood smears in the canine blood. Blood...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • Journal of clinical pathology

دوره 43 8  شماره 

صفحات  -

تاریخ انتشار 1990