Clin. Lab. 2/2024 433 Clin. Lab. 2024;70:433-436 ©Copyright SHORT COMMUNICATION Performance Evaluation of the Atellica Hema 580 Automated Slidemaker and Stainer for Morphology Review Elise Schapkaitz 1, 2, Gaone A. Moalosi 1, 2, Anima Baiden 1, 2, Shilla Raburabu 2 1 Department of Molecular Medicine and Hematology, University of Witwatersrand Medical School, Johannesburg, South Africa 2 Department of Hematology, National Health Laboratory Service, Johannesburg, South Africa SUMMARY Background: Improved measurement technologies for automated peripheral blood smear (PBS) preparation have led to a more efficient workflow. In this report, we evaluated the Atellica Hema 580 automated slidemaker/stainer (Siemens Healthineers, Tarrytown, NY, USA). Methods: The quality of PBS prepared and stained on the Atellica Hema 580 slidemaker/stainer were compared with manual films stained on the Hematek 3000. A 200-cell differential and morphology assessment were perform- ed by two competent morphologists. Results: Macroscopically, the automated PBS were of excellent quality, with a gradual transition in thickness, smooth edges, and sufficient length. The morphology was comparable between manual and automated PBS. Cor- relations between the manual and automated PBS were 0.95 (95% confidence interval: 0.92 to 0.96) for neutro- phils, 0.63 (95% CI: 0.50 to 0.74) for monocytes and 0.90 (95% CI: 0.85 to 0.93) for lymphocytes. Conclusions: The performance of Atellica Hema 580 automated slidemaker/stainer was comparable to the manual method and represents a reliable automated solution for medium to large sized laboratories. (Clin. Lab. 2024;70:433-436. DOI: 10.7754/Clin.Lab.2023.230818) Correspondence: Elise Schapkaitz P.O. Box 28985 Sandringham 2131 South Africa Phone: +27 824592238 Email: elise.schapkaitz@nhls.ac.za _______________________________________________________ Short Communication accepted August 29, 2023 KEYWORDS validation, automated slidemaker/stainer, quality, Atellica Hema INTRODUCTION Atellica HEMA 580 Analyzer (Siemens Healthineers, Tarrytown, NY, USA). This multi-parameter automated analyzer performs a FBC and DIFF by impedance varia- tion and fluorescence flow cytometry. The system is composed of an analyzer, a compressor unit (Atellica data manager: external data management) and an auto- mated slidemaker stainer. Peripheral blood smears (PBS) are prepared according to the hematocrit and use the May-Grunwald Giemsa or modified Wright staining The full blood count (FBC) and differential (DIFF) is one of the most important diagnostic and screening lab- oratory tests. Recently, Siemens Healthineers launched a new generation protocol. The slidemaker stainer uses the wedge method which consists of adjustable settings E. Schapkaitz et al. Clin. Lab. 2/2024 434 for sample volume, speed, and angle of the wedge [1]. This has been shown to improve the consistency of the cell distribution and thereby the precision of the DIFF count [2]. Automated staining offers several advantages including efficient workflow, shorter staining times, and enhanced sample throughput. The Atellica HEMA 580 has a throughput of 120 slides per hour. Nonetheless, only a few studies have evaluated the quality and per- formance characteristics of automated slide makers and stainers [2-4]. Therefore, this study evaluated the per- formance of the Atellica Hema 580 automated slide- maker stainer compared to reference manual PBS. MATERIALS AND METHODS The automated slidemaker stainer, which is 91 cm wide x 72 cm deep x 74 cm high was installed at the Char- lotte Maxeke Johannesburg Academic Hospital, Johan- nesburg, South Africa (Table 1). Daily quality control and maintenance were performed according to the man- ufacturer’s specifications. A total of 105 patient samples were selected from the routine workload between April and May 2023. These samples included normal and ab- normal hematology profiles representative of the patient population. Samples were collected in K2EDTA tubes (Vacutainer®, Becton Dickinson, Plymouth, UK) for analysis. PBS were prepared and stained on the auto- mated slidemaker stainer using the Wright-Giemsa stain and compared to manual PBS, stained with Wright-Gi- emsa Stain on the Hematek 3000 (Siemens Healthine- ers, Tarrytown, NY, USA). The quality of PBS prepared and stained on the Atellica Hema 580 automated slidemaker stainer were evaluated and graded as excellent; good; adequate for diagnosis, suboptimal or poor. A 200-cell DIFF was performed, using light microscopy equipped with 50 x and 100 x objectives, in the monolayer of the automated and ma- nually prepared PBS of each sample. This was perform- ed by two competent morphologists according to the Clinical and Laboratory Standards Institute H20-A2 verification protocol [5]. Others included immature granulocytes, blasts, abnormal lymphocytes, plasma cells, nucleated red blood cells. The 95% confidence in- tervals for imprecision of the DIFF counts were derived from the standard error of proportion calculations. Red blood cell (RBC) morphological assessment was per- formed in terms of size, shape, and pallor based on the International Council for Standardization in Hematolo- gy recommendations [6]. This study was approved by the Human Research Ethics Committee of the Universi- ty of the Witwatersrand (M090688). RESULTS AND DISCUSSION Macroscopically, the automated PBS were of excellent quality for morphological assessment, with a gradual transition in thickness, smooth edges, and sufficient length (Table 2). Stain artefact and precipitant were ob- served in 9 (8.6%) automated PBS; however, all PBS were judged acceptable for morphological assessment. Morphological assessment revealed that the erythrocyte stain and morphology of RBC were comparable be- tween manual and automated PBS. In addition, the qual- ity of the platelet stain was comparable between the manual and automated PBS. The distribution of white blood cells (WBC) within the monolayer were compara- ble between the manual and automated normal and ab- normal PBS. A ‘line’ of cells with increased WBC clumps was observed at the tail of most of the automat- ed PBS. This morphologically significant finding has been described with other automated slide-makers. Pre- viously, Simson et al. [3], observed a ‘line’ of cells with increased WBC clumps in 16.5% of the 139 samples ex- amined on the Beckman Coulter LH755 automated slide-maker (Beckman Coulter Inc., Miami, FL, USA) [3]. Nonetheless, consistent with our findings, there were no significant differences in the distribution of WBC types and DIFF counts between the manual and automated PBS. In the analysis of the 200-cell DIFF counts, correlations in the neutrophil, monocytes, lym- phocyte, eosinophil, other and nucleated red blood cell percentages between the manual and automated smears were very strong/strong (Figure 1). Basophils, however, showed a poor correlation as these cells accounted for a very low proportion of the total WBC. CONCLUSION The performance of Atellica Hema 580 automated slidemaker stainer was comparable to the manual meth- od. This automated slidemaking and staining technolo- gy provided improved workflow efficiency, standard- ization, and stain reliability. This represents a reliable automated solution for medium to large sized laborato- ries. Acknowledgment: The authors thank Mrs. Judy Moodley from Siemens Healthineers for her assistance with this study. We would like to thank Mr. Devind Peter of the Faculty of Health Sciences Library for his assistance with retrieval of full-text references. Declaration of Interest: The author(s) declare no conflicts of interest with re- spect to the authorship and/or publication of this article. Verification of an Automated Slidemaker/Stainer Clin. Lab. 2/2024 435 Table 1. Atellica Hema 580 slidemaker and stainer setup. Specifications Maintenance Input-160 slides with continuous feeding capability Drop volume < 5 µL Wedge landing angle on the slide 40 - 45 Wedge stays in contact with the drop before making the smear for < 2,000 ms Smear speed < 5,000 steps/s Daily start-up Weekly shutdown Glass particles tray cleaned weekly Staining wells cleaned monthly Staining plate cleaned monthly Replace stain and alcohol air valves six-monthly Frosted, rounded corner, beveled edges, 1.0 mm thick glass slides (FF-100, Matsunami, Osaka, Japan). Table 2. Quality assessment for slidemaker and stainer. Parameters Atellica automated method Reference manual method Approximately 2.5 cm long, terminating 1 cm from the edge excellent excellent Narrower than the width of the slide with smooth edges excellent excellent The gradual transition from thick to thin areas ending in a feather edge excellent good No staining artefacts/stain precipitate adequate excellent Sufficient working area with evenly distributed neutrophils, monocytes, and lymphocytes excellent good The side edges should contain less than 2 to 3 times the number of cells per 100 x field than the body of the film adequate excellent Except for certain forms associated with pathological states less than 2% of the white blood cells should be disrupted or unidentifiable good excellent Nuclei and cytoplasm of different cell types stained satisfactorily excellent good Neutrophil % Lymphocyte % 0.95, 95% confidence interval 0.92 to 0.96 0.90, 95% confidence interval 0.85 to 0.93 E. Schapkaitz et al. Clin. Lab. 2/2024 436 Monocyte % 0.63, 95% confidence interval 0.50 to 0.74 Figure 1. Correlation of the neutrophil, lymphocyte, and monocyte percentages between the manual and automated peripheral blood smears. References: 1. Atellica-Hema 580 Analyzer operators guide. Tarrytown, NY: Siemens Healthineers; 2021. https://www.siemens-healthineers.com/hematology/systems/atel lica-hema-570-580-analyzers 2. Pawlick G, Relopez J. Kaiser Permanente interlaboratory abnor- mal cell study comparing slide quality of the Sysmex SP-100 au- tomated slide preparation unit to manual technique. Sysmex Journal International. 2000;10:26-9. https://www.sysmex.co.jp/en/products_solutions/library/journal/v ol10_no1/summary04.html 3. Simson E, Gascon-Lema MG, Brown DL. Performance of auto- mated slidemakers and stainers in a working laboratory environ- ment - routine operation and quality control. Int J Lab Hematol 2010 Feb;32(1 Pt 1):e64-76. (PMID: 19220552) 4. Kwak MS, Jeong IH, Cho SS, Woo KS, Han JY. Performance Evaluation of the Abbott Alinity hs Blood Slide Maker/Stainer. Ann Lab Med 2022 Jul 1;42(4):482-4. (PMID: 35177570) 5. Clinical Laboratory Standards Institute. H20-A2: Reference Leu- kocyte (WBC) Differential Count (Proportional) and Evaluation of Instrumental Methods, 2nd edn. Wayne, PA: Clinical Labora- tory Standards Institute; 2007. https://clsi.org/media/1400/h20a2_sample.pdf 6. International Council for Standardization in Hematology, Writing Group; Briggs C, Culp N, Davis B, d'Onofrio G, Zini G, Machin SJ; International Council for Standardization of Hematology. ICSH guidelines for the evaluation of blood cell analysers includ- ing those used for differential leucocyte and reticulocyte count- ing. Int J Lab Hematol 2014 Dec;36(6):613-27. (PMID: 24666725) https://www.siemens-healthineers.com/hematology/systems/atel%20lica-hema-570-580-analyzers https://www.siemens-healthineers.com/hematology/systems/atel%20lica-hema-570-580-analyzers https://www.sysmex.co.jp/en/products_solutions/library/journal/vol10_no1/summary04.html https://www.sysmex.co.jp/en/products_solutions/library/journal/vol10_no1/summary04.html https://clsi.org/media/1400/h20a2_sample.pdf