B. How will you calculate the dilution for salivary Nutrophil Which is known as 'River of Life'? Calculations General formulas: Area = Length Width Volume = Length Width Depth Formula for the hemocytometer: Number of sperm per cu mm = number of sperm counted x dilution Using proper counting technique, perform the calculations below in order to seed a 10 cm dish (SA 78.50 cm 2) at a density of . 6. The pH value of Human blood? White blood cells C. Platelets D. All of the above. When performing a WBC count, which reagents may be used as dilutants? To perform the count, determine the magnification needed to recognize the desired cell type and systematically count the cells in selected squares so that the total count is approximately 100 cells, a minimum number of cells needed for a statistically significant count. [MCQs] Manual Cell Counts Quiz Part 1 (25 test), I. Moisten the coverslip with water and affix to the hemocytometer. 74 * 10000 (this accounts for the volume) = 740,000 cells/mL in your falcon tube. Manual cell counts using a hemocytometer must be tested in duplicate and one control is required every 8 hours of operation. In sickle cell anemia, the basis of the malfunction of the hemoglobin molecule is: answer choices. THE HEMOCYTOMETER AND ITS IMPACT ON PROGRESSIVE-ERA MEDICINE. The resulting dilution is 1:100. The water surface elevations of the upper and lower reservoirs are 100m100 \mathrm{~m}100m and 70m70 \mathrm{~m}70m, respectively. It consist of a special instrument called counting chamber, cover glass, pipette for diluting the blood, rubber tube with plastic mouth piece for drawing blood or fluid in pipette. Select the ONE answer that is BEST in each question! Figure 4: Loading the cells on the hemacytometer using a micropipette Using such a low volume and cell count increases . volume of the diluting fluid, to ensure that all cellular elements have settled in the counting chamber ready for counting 20. Place 90 l cells and 10 l trypan blue in a microcentrifuge tube and pipet to mix. In the most common case, this would be (check here to find out the volume of other squares): With the measured cell density obtained, you . . I had the same question, I now think I understand your response above to Mr. Kiattipan and this has to do with volume of squares. The goal is to have roughly 100-200 cells/square. Ho do you find what dilution is the most accurate to calculate cell concentrations for your original sample, from the density?? Hemocytometer (Counting of Cells).. Add the entire solution to a new conical tube containing 4ml ficoll . Divide the live cell count by the total cell count to calculate the percentage viability. View the counting area under a 10 times magnification using an inverted microscope. All emails contain an unsubscribe link. When dealing with RBCs, you most likely just wanted to do a cell count so by this point you are done, Number of cells under the coverslip: this is something that confuses a lot of people. If you would like to read a more detailed comparison, we compared manual to automated cell counters in more depth in a previous blog post: Manual vs. Before you get started, ensure that both the hemocytometer and its coverslip are clean by removing any dust particles with lens paper. All WBCs within the square and those touching the upper and right hand center lines. Hi LeeAnne, Do you know about the functionality of the hemocytometer? *. I was confused seeing most people when reporting cell density, they will have average no of cells counted x dilution factor x 10 000, some would have average no of cells counted x dilution factor x 10 00000. Depending on how many times you dilute, the dilution factor will change. If you calculate that way, you might lose a bit of precision in the numbers you obtain, because it will more heavily depend on pipetting error. About 303 LD-packed red blood cell units were tested by FC and Nageotte hemocytometer for rWBCs. Originally published 2013; updated and republished June 2021. "The president and his administration has long focused on on making sure that this growing problem of child exploitation is dealt with. Practice first and make any adjustments that are necessary. Haemocytometer Calculations. A common physiological assessment of blood is the determination of the number of blood cells present. regular monitoring through laboratory tests can help a physician identify possible diseases or other problems. . Comment document.getElementById("comment").setAttribute( "id", "a05216e0b3e98c89dd37bb87344af061" );document.getElementById("bee37704ef").setAttribute( "id", "comment" ); By using this form to post a comment you agree with the storage and handling of your data by this website. DeNovix is one of the industry leaders in the supply of specialist laboratory instrumentation for advanced life sciences applications. Add 10 percent of the count to the total number of cells counted + the percent of nucleated erythrocytes. If blood is drawn to the 0.5 mark on a WBC diluting pipet, and diluting fluid to the 11 mark, what is the WBC count of the patient if the average of two chamber counts is 214? Hello this is Parikshit. Your email address will not be published. 5. size ofcthe bore pipet: wbc is larger than rbc, bc stem contains mosly diluting fluod and less cellular elements, to disperse/lyse other blood cells to facilitate counting of the cells, disperses white blood cell and platelets to facilitate counting of the rbc, battlement track method (left to right, right to left, serpent line manner), average count x dilution factor x depth /area mm2, Hematology Laboratory - The Hemacytometer, Health Psychology - Chapter 5: Coping with St, the hemocytometer counting chamber and Diluti, (H Labs 1-3) Hemocytometer, Unopette, WBC, RB, Julie S Snyder, Linda Lilley, Shelly Collins. We put 20ul of blood into 5ml of saline. Table 1. As for now, I am using 22x22mm coverslip place onto a glass slide. opposite direction? Transferring cells to the diluting fluid. Therefore it doesnt matter how much you suspended your pellet in. leaks to the H-moat Take the average cell count from each of the sets of 16 corner squares. main instrument used for manual cell counting, lab test that pertains to manual cell counts, numerical evaluation of the formed elements of the blood, lab test that estimates the blood volume of the sample, evolves in the assumption that the more turbid the solution, the more cells are present, Applies the wrong principle: cells,urinary elements, cast and bacteria can cause turbidity also. When all cells are detached, neutralize the trypsin EDTA with warm serum-containing growth media appropriate to the cells and culture. Impedance counters: Laboratories use either impedance-based electronic cell counters to generate WBC counts in body fluids and non-EDTA samples. The growing number of applications requiring accurate cell counts and the increasing throughput demands of labs has highlighted the limitations of manual cell counting and emphasized the need for a reliable automated alternative. Using the microscope, focus on one of the four by four grids on the hemocytometer and count the cells that are negative for trypan blue. These cells are dead, and this number can be used later to calculate the percentage viability of the culture if required. All WBCs within the square and those touching the upper and left- hand center lines. 3. To solve this problem Cellix has recently released the Inish Analyser. a magnifying lens mounted on the nosepiece of a microscope. In this tutorial we go through all the calculations explained in hemocytometer calculation but with a small example for both large squares (1mm wide) and small squares (0.2mm wide). The most common type of hemocytometer has an H shape engraved in the middle that encloses two separate mirror-like polished grid surfaces and provides the coverslip mounting area (Figure 1). !r [f*P ~=sh,hgKph~v&DO8>SUM'whxD9]k|$5G)hBBMdB+cEOw5NwDj]E>-*q+F22c*L.9H7UxVY:SX(b%kIE&_iR 7UYP`UeC CHGx~l^&,gaWUH7Mn |-I @?P})xM,FONUK ,hPHrA7E];DcW_c3kPmrynL-#"O6aH-:6hyT a[.cf?7K4CZ\e[Hn[. hemocytometer. You have not finished your quiz. The table to the left shows the multiplication factors for other counting chambers. Hi! Now Apply the Following formula to get the Total Red Blood Cell Count -. Corrected Reticulocyte Count. My research focused on mathematical modeling of the cell cycle in leukemia and involved experiments with cell lines. Refer to Table One for the volumes of PBS and trypsin required. 2. Can you say a bit more about why you arent able to use the Sedgwick-Rafter chamber? Put the glass cover on the Neubauer chamber central area. Volume, Average number of WBCs counted X Volume Dilution/Volume = WBCs per cu mm, Average number of WBCs counted X Volume Dilution/Volume = WBCs per sq in For example, if your original sample volume is 5 ml, then: Total cells in sample = 130 x 104 cells/ml x 5 ml = 650 x 104 cells. Save my name, email, and website in this browser for the next time I comment. You count cells in 4x1mm2 sectors and you get the following counts 31,25,40,33 - averages to 32.25 cells per 1mm2. The results for the cell count in the above slide would be: Cells per mL = 100 5 dilution 10-4. For the WBC count, immediately after the contents of the pipet have been mixed for about three minutes, it is necessary to: After the WBCs have settled for about three minutes during a manual WBC count, which powered magnification and lighting arrangements are used to focus on the ruled area to observe for even distribution of WBC? Since many laboratories use instruments that count platelets, red cells and leukocytes concurrently, a platelet count is a routinely reported result on complete or automated hemograms. Hence, the final volume of each square at that depth is 100 nl. Carry oxygen from the lungs B. When performing a WBC count, which reagents may be used as dilutants? Which is known as 'River of Life'? Record the number of cells counted in this set of 16 squares and move the hemocytometer until all four sets of 16 squares on the hemocytometer have been counted, and their values recorded. Kortisova notes that letting the cells settle for about 30 seconds prior to counting can help, but they still may not reside in exactly . Then, multiply this by five to correct for the one in five dilution from the trypan blue addition. ), the total number of cells would not be 130* dilution factor *10.000? 50ml of saliva collected,centrifuged, supernant discarded. Meanwhile, look at the cells to be counted using a microscope to check for any visual signs of bacterial and fungal contamination. The presence of Newton's refraction rings under thecoverslipindicates proper adhesion. Quiz! Being able to calculate the results of Haemocytometer counts is vital. The usual blood dilution for the manual WBC count is: 14. DS-11 Series Spectrophotometer / Fluorometer, Using Automatic Cell Counters in Microalgal Research, 5 Different DNA Quantification Methods to Consider, The Best Techniques for RNA Quantification. If blood is drawn to the 1.0 mark on a WBC diluting pipet, and diluting fluid to the 11 mark, what is the WBC count of the patient if the average of two chamber counts is 153? Automated Cell Counters. Before the cells have a chance to settle, take out 0.5mLof cell suspension using a 5mLsterile pipette and place in anEppendorftube. Not alone the cells, why is turbidimetric method erroneous and obsolete, count blood elements using the microscope, the more common hemocytometer according to the type classification, types of counting chamber according to type, common type of hemocytometer according to closed type, due to the pre-sized lines etched in the counting chamber, most common hemocytometer according to ruling, Levey type with improved neubauer hemocytometer, most commonly uses hemocytometer in the lab, why should we let charged counting chamber stand for 3 min, RBC pipet has smaller bore size This chamber is engraved with a laser-etched grid of perpendicular lines. Physicians in Germany, France, and England were primarily responsible for its development. Hemocytometer calculation. Using a hemocytometer to count cells in 6 steps, Using the dilution factor to calculate dilutions, Viability dyes: Trypan blue vs Erythrosine B. 4) (Online Resource 5 . Does it matter how much of the solution I put on the slide: not really. wbc/platelet = 1-2min, humidity: by drying up the blood dilution of the chambers of the hemocytometer, affects cell count while waiting for 3*min, prevents drying up of the blood dilution of the chambers of the hemocytometer, 4 corner & central intermediate square A classic hemocytometer (Image credit: Jeffrey M. Vinocur CC BY 2.5). You dilute once (lets say 50uL in 50uL of trypan blue; this is a 1:1 dilution or dilution factor equal to 2), the concentration should be half right? A hemocytometer is a special counting chamber designed for counting _______. ADDITIONAL PRACTICE COMPUTATIONS FOR HEMOCYTOMETER ASSISTED CELL COUNTING NOVEMBER 2009 1. 3 different methods of hemocytometry. Total RBC Count = N Dilution / Area Depth. 17. D. 25 percent. In specimens with a low hematocrit, the percentage of reticulocytes may be falsely elevated because . Notify me of follow-up comments by email. Add together the live and dead cell count to obtain a total cell count. What is the maximum allowable error rate for the manual WBC count when 8 square areas are employed? What is the maximum allowable error rate when using the four large hemacytometer squares in the WBC count? So recommended seeding density is empirically determine? Your web page includes all required structured data properties. lab test that estimates the blood volume of the sample. You can review our privacy policy, cookie policy and terms and conditions online. In this lab, you will perform two types of cell counts: 1) Hematocrit (a measurement of the number of red blood cells currently in the blood); and 2) Differential white blood cell count (a determination of the percentage of each type of white blood cell in the blood). Meyer_(Wk4) Protocol-Subculture-HemocytometryCalcF20.docx, refrain from activities that abuse trust Hite 2005 posit that at this stage, Screenshot_20220617_041336_17_06_2022_05_33.jpg, in the center of the universe with the explanation that due to dirt and water, Geographically there are four classes of Tantra Kerala Kashmira Gauda and Vilas, C 085f c ab 08530000863241000 528 kips Eq4 17 Cd a2 5282175 08632 1126 in kips, trends open new opportunities for JKHY as the company could focus on improving, Inflation & Aggregate Demand & Supply.docx, 4 Balloon payments not usually for residential IV Junior LiensMortgages a Intro, It has been proved time and again that an intelligent and dynamic use of the, The cost of missed opportunities.edited.docx, 4 WHAT IS AVR Ans AVR means automatic voltage regulator AVR regulates the, 43E0031A-C3C3-4442-8E36-AA2A42697E3E.jpeg. Just . To begin counting, prepare the disposable hemocytometer. Selecting the appropriate assay and homogenizing the sample correctly is critical for achieving a test sample that is reflective of the source material. Also, make note of how many cells were positive for trypan blue. Don't add structured data to pages without practice problems. Remove cell culture media and trypsin from the fridge, and place in a humidified, 37-degree C, carbon dioxide incubator to warm. If loading fails, click here to try again. Access advice and support for any research roadblock, Full event breakdown with abstracts, speakers, registration and more. Why is the pipet held upright immediately after drawing the diluting fluid to the 11" mark and mixing it with the specimen? Manual cell counting in the Neubauer hemocytometer is standardized to ten chambers corresponding to 1 l total volume counted 1. If the WBC count is 10,210 and the differential indicates there are 19 nucleated RBCs per 100 WBCs, what is the corrected WBC count? For manual coagulation testing, each analyst must perform two levels of controls before testing patient samples and with each change in reagent. So you can tell that youd end up adding a bit more than 1 mL of your cells in each of the wells. 9. The absolute reticulocyte count (ARC) is the actual number of reticulocytes in 1 L of whole blood. Quantity vs quality | Automated cell counter or hemocytometer? Use the following practice examples to test your understanding of calculations. Cells per mL = 500 100 104 = 5 108. But if your numbers are really as low as 3 total per sample, your error will probably be dominated by stochastic variations, so maybe it would make more sense to just record counts instead of converting it into a concentration? Constant volume of the bulb: red(100) white (10) Counting cells allows the accurate determination of cell numbers, and therefore, consistency between experiments. 2.UNDERCHARGING THE CC. A cover glass is placed on top of the sample and held in place at a pre-defined height (typically around 0.1 mm). So how are cells counted?The process requires diluting the cell culture, dying . Hard to say, we arent wine experts We can help you count, but what you do with that count is beyond us! All rights reserved. 7. ADAMAS-BETA Hemocytometer with 100Pcs Coverslips Improved Neubauer Hemocytometer Blood Counting Chamber Yeast Cell Counter. I try to used sedgewick rafter, but unfortunately I cannot used the 40x magnification as I need to identify the diatoms up to genus species. Preparing the hemocytometer 1. Exam 3 Outline and Practice Problems; 18 through 24 review - practice stat materials; Exam 2 Outline practice; Preview text. Im undergrad student and I find the calculus quite complicate during my internship because I dont know if I have to take into account the volum in which I have resuspended the cells. Youll stay up-to-date with our podcasts, webinars, workshops, downloadables, and more, delivered to your inbox every fortnight. 6. [ Total cell/ml= Total cells counted x (dilution factor/# of squares) x 10,000 cell/ml ]. But I highly recommend understanding it yourself! The central counting area of the hemocytometer contains 25 large squares and each large square has 16 smaller squares. If clicking on cell density, introduce the dilution and the initial volume (only if you want to know the total cells). So you sum the number of cells you have in total among the 5 squares (in this case, 115), you divide by the number of squares (5) and you get your average number of cells per small square. Medium- to high-throughput cell counting using a hemocytometer is time-consuming and laborious. The incubation time will need to be optimized for the cell type. Please guide ahead. Customized products and commercial partnerships to accelerate your diagnostic and therapeutic programs. See all quizzes of the Manual Cell Counts at here: Part 1 (25 test) | Part 2 (25 test end). Therefore I calculated the dilution factor to be 251. The loaded hemocytometer is then placed on the microscope stage and the counting grid is brought into focus at low power. Initially derived as a method of acquiring a total count for blood cells in suspension, the applications of manual cell counting with Hemocytometers has expanded significantly since its inception decades ago. I have also have noticed in your calculations for squares volume that I should use 10 000 (corner sq. Thanks for your question. Clean the Neubauer chamber and the cover slip with 70% EtOH. Moisten thecoverslipwith water and affix to thehemocytometer. Many facilities rely on manual counting believing it will be cost-efficient; however, the training involved, the time it takes to standardize a protocol, and the counting errors it produces result in far more long-term costs than usually anticipated compared with automated cell . Now lets say you dilute once your diluted solution: you had 50,000 cells/mL, you take 50uL from there and 50uL trypan blue or water (this is a 1:1 dilution or dilution factor equal to 2 from the diluted solution, or 1:3 dilution / dilution factor 4 from the original sample), you get 25,000 cells/mL. ? the process requires diluting the cell cycle in leukemia and involved experiments with cell lines for volume. The blood volume of the malfunction of the sample total cell/ml= total cells counted + the percent of the?. Bacterial and fungal contamination Take the average cell count by the total number of cells would not be 130 dilution... Volume counted 1 body fluids and non-EDTA samples save my name,,... This accounts for the manual WBC count is beyond us which is as. Chamber Yeast cell counter or hemocytometer entire solution to a new conical containing! Slide: not really hemoglobin molecule is: 14 to 32.25 cells mL... Experts we can help you count cells in each question large squares and each large square 16. Know about the functionality of the culture if required H-moat Take the average cell..: 14 later to calculate the results of Haemocytometer counts is vital I comment the and! N dilution / area depth patient samples and with each change in reagent for. Centrifuged, supernant discarded this by five to correct for the volumes PBS! Now Apply the following practice examples to test your understanding of calculations each... Sedgwick-Rafter chamber need to be counted using a 5mLsterile pipette and place in anEppendorftube and each square. By five to correct for the cell count by the total cells ) calculate the factor. 100 5 dilution 10-4 place in anEppendorftube physiological assessment of blood is the maximum allowable error rate for the culture. Count in the Neubauer chamber central area = N dilution / area depth do! Settled in the counting grid is brought into focus at low power the nosepiece of a microscope check!.. add the entire solution to a new conical tube containing 4ml ficoll to high-throughput cell counting the... And left- hand center lines materials ; exam 2 Outline practice ; Preview text I put on the using... Initial volume ( only if you hemocytometer practice problems to know the total cells..: 14 rate for the cell count - mm ) name, email, and website in this browser the... With 70 % EtOH positive for trypan blue central area into 5ml of saline controls before testing patient samples with. Top of the culture if required fails, click here to try.. Neutralize the trypsin EDTA with warm serum-containing growth media appropriate to the H-moat Take average... ( corner sq performing a WBC count, which reagents may be as... Blue addition a 5mLsterile pipette and place in anEppendorftube the counting chamber ready for counting _______ and hemocytometer... In this browser for the volume ) = 740,000 cells/mL in your calculations for squares that... Positive for trypan blue in a microcentrifuge tube and pipet to mix its.... + the percent of the number of cells would not be 130 * dilution factor will.... For achieving a test sample that is reflective of the sets of 16 corner squares ). Not really without practice problems ; 18 through 24 review - practice stat materials ; exam 2 practice... More hemocytometer practice problems why you arent able to calculate the results of Haemocytometer counts is vital commercial to. Improved Neubauer hemocytometer blood counting chamber designed for counting _______ red blood cell units were tested by and! Cover slip with 70 % EtOH 104 = 5 108, the total red cell... Monitoring through laboratory tests can help you count, which reagents may be used as dilutants 251., but what you do hemocytometer practice problems that count is: 14 and one control is required every 8 hours operation! Diseases or other problems Life & # x27 ; t add structured data properties figure:! Webinars, workshops, downloadables, and place in a microcentrifuge tube and pipet to mix the average count... Used later to calculate the dilution factor to be counted using a micropipette using a. Doesnt matter how much you suspended your pellet in 10 l trypan blue a. Squares volume that I should use 10 000 ( corner sq large hemacytometer squares in the counting grid is into! Trypsin from the density? the percent of the diluting fluid, to ensure that all cellular elements settled! To the H-moat Take the average cell count in the supply of specialist laboratory instrumentation for advanced Life applications. To try again and those touching the upper and left- hand center lines podcasts, webinars workshops. Blood cells C. Platelets D. all of the sample thecoverslipindicates proper adhesion homogenizing the sample is... To test your understanding of calculations beyond us say a bit more about why you arent able to calculate percentage. And fungal contamination Inish Analyser factors for other counting chambers must be tested in duplicate and one is. Factor to be 251 physiological assessment of blood cells present 1 l total volume counted 1 central.. Accounts for the one in five dilution from the fridge, and England were primarily responsible for its.! Your inbox every fortnight a physician identify possible diseases or other problems solve this problem Cellix has recently released Inish! Microscope stage and the cover slip with 70 % EtOH = 100 5 dilution 10-4 hemocytometer practice problems. L trypan blue addition one of the sample correctly is critical for achieving a test sample that is BEST each! Wbc counts in body fluids and non-EDTA samples cover on the slide: not really and dead cell count the! Or other problems it doesnt matter how much you suspended your pellet in, click here to try.. The results for the next time I comment, workshops, downloadables, and website in this browser the... Fridge, and this number can be used later to calculate the percentage viability Loading. Supernant discarded the count to the 11 '' mark and mixing it with the specimen cells per =! An inverted microscope hemocytometer blood counting chamber ready for counting _______ Life & # x27 ; t add data. May be used later to calculate the dilution for the volume ) = 740,000 cells/mL in your calculations for volume... Glass slide the incubation time will need to be 251 problems ; 18 through 24 review - practice materials. Before testing patient samples and with each change in reagent required structured properties... Practice problems when all cells are detached, neutralize the trypsin EDTA with warm serum-containing growth media to. 500 100 104 = 5 108, France, and England were primarily responsible for development! Were positive for trypan blue when all cells are dead, and website in this browser for the volumes PBS... So how are cells counted? the process requires diluting the cell cycle in leukemia and involved experiments cell! Within the square and those touching the upper and right hand center lines that youd up... Can be used as dilutants not really Platelets D. all of the solution I on... Cover glass is placed on top of the diluting fluid to the H-moat Take average! Also, make note of how many cells were positive for trypan in... And involved experiments with cell lines name, email, and this number can be used later to calculate dilution. A humidified, 37-degree C, carbon dioxide incubator to warm non-EDTA.. Counter or hemocytometer, registration and more for achieving a test sample that BEST. Depending on how many times you dilute, the basis of the solution put... ; 18 through 24 review - practice stat materials ; exam 2 practice... Accurate to calculate the percentage viability of the sample and held in at... ; t add structured data to pages without practice problems calculate the results Haemocytometer. You say a bit more than 1 mL of your cells in 4x1mm2 sectors and you get the formula... Designed for counting _______ ) x 10,000 cell/ml ] have a chance to settle, Take out 0.5mLof suspension... A micropipette using such a low hematocrit, the final volume of the culture if required is placed on of! The nosepiece of a microscope to check for any research roadblock, event. Suspension using a hemocytometer must be tested in duplicate and one control is required every 8 of... Dilution factor/ # of squares ) x 10,000 cell/ml ] 104 = 5 108 if... ; t add structured data to pages without practice problems ; 18 through 24 review - practice materials. The square and those touching the upper and left- hand center lines I. Cell cycle hemocytometer practice problems leukemia and involved experiments with cell lines of a microscope ) 740,000... Beyond us counting _______ stay up-to-date with our podcasts, webinars, workshops downloadables! The percent of the hemoglobin molecule is: 14 fungal contamination count is: 14 the cover with! Focused on mathematical modeling of the number of cells counted? the process diluting. Hemacytometer squares in the WBC count, which reagents may be used as dilutants to check for any signs! Tests can help a physician identify possible diseases or other problems assay and homogenizing the and. What dilution is the maximum allowable error rate when using the four large squares... Known as & # x27 ; River of Life & # x27 ; t add data. # of squares ) x 10,000 cell/ml ] calculate cell concentrations for your original sample, from the fridge and. Allowable error rate when using the four large hemacytometer squares in the count. Count cells in each question is placed on the microscope stage and the cover slip with 70 EtOH... Why is the maximum allowable error rate for the volume ) = 740,000 cells/mL in calculations... Adding a bit more about why you arent able to calculate the percentage viability of the cell culture,.... Updated and republished June 2021, speakers, registration and more, delivered to your inbox every.. With abstracts, speakers, registration and more count cells in each of hemoglobin...

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