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Hæmatological Technique. Part VIII

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Hæmatological Technique. Part VIII Special Articles be -dried HEMATOLOGICAL technique spread; they should quickly by waving them to and fro under a fan (vide part VI). Part VIII Staining.?Combined staining with May- Griinwald and Giemsa stains gives the most (Lond.) By L. EVERARD NAPIER, f.b.c.p. satisfactory result?the nuclei stain deeply, the and connecting chromatin filaments are well shown, C. R. DAS GUPTA, Mi. (CaU. , while the granules also show characteristic ,n D-^'.Calcutta, and size. If and Giemsa (From the School of Tropical Medicine, colour May-Griinwald stains are not with counts available, proper staining The Arneth and Schilling or Leishman's not as K (13) Wright's stains, though the satisfactory, gives quite good results. Iron- or Arneth (1904) divided ? aCCOrding for these main c g1 hamiatoxylin haematoxylin staining into five iej which counts in to of the Romanowski ^orphonuclears m the preference any t? the number of is segments into groUps stains sometimes advocated; while these ln were the their turn classified -? TTnder stains out the nuclei and the chromatin of the lo) bring according to the shape c0m- filaments the colour of the is not the count in well, granules ?riginal classification i-cati0n distinctive so that differentiation between Phcated and had only limited PP, and (1927) neutrophils and eosinophils is very difficult. general practice. Cooke lint taking the Arne and simplified, divisions .4. Arneth count _ y^odifiedlnto account the five original into the only going The nucleus the Arneth count without of the neutrophil polymorpho- ?f be lntncate subdivisions of Arneth. ,vy,\ch the nuclear cells is divided into lobes which may * or of devised a nuclear separate joined together by fine filaments Schilling < - iinpie differential countmfetk?f and chromatin. Cooke's criterion of separate lobu- leucocyte Jpnroorated be l?be count of the'neutrophils.were lation is that either the lobes should distinctly and form of a ^c?rPO^ or should be a fine considered in the '? separate, joined together by but not broad bands of -Apparatus required (vide part Sctaiua0hilling B chromatin filament, by the Arneth nuclear material. Lobulation of the nucleus, it Smears.?Smears for andyd C0llnts should be very thin, 98 THE INDIAN MEDICAL GAZETTE [Feb., 1941 an is assumed, gives indication of the age of the & cell?the less the lobulation the is the class V by 5, and then adding together younger of ce cell. The youngest cell in the normal peripheral dividing the sum total by the number counted. circulation is one which has a nucleus shaped ^ ^ The means of the nuclear lobes like the letter C, while the cells with five or weighted these three are calculated in more distinct nuclei are the oldest. The prin- examples ciple of the method is to make a differential following way :? count of the to group the cells neutrophil cells, 3 according to the lobulation of the nuclei, and to Example 1 Example 2 Example calculate the percentage of cells in each group. The cells of the various types can be described IX 10 = 10 X 41 = 41 X 9 = = I as follows :? 2 X 25 = 50 X 44= 88 X 22 = 3 X 47 = 141 X 14 = 42 X 36 Class I. in which ? lg& Neutrophil granulocytes 4 X 16 = 64 X 1 = 4 X 23 one or = 50 the nucleus has only definite lobe, two 5 X 2 = 10 X 0 = 0 X 10 or more lobes joined by a definite band of I chromatin and not by thin filaments. 100 275 100 175 100 301 Class II. When there are two separate lobes joined by one thin filament, or when there are = 3.0* two completely separate lobes. Weighted mean 2.75 1.75 Class III. When there are three separate & lobes connected by two thin filaments, or there Interpretation.?Immature granulocytes l are three completely separate lobes. cells of Arneth class I and some of class II) R, , Class IV. When there are four separate none of class III, IV or Y, are found in ^ I lobes connected by three thin filaments, or there bone marrow. If there be a sudden stimulus the bone marrow a<3 are four completely separate lobes. to produce granulocytes, ^ j an a Class V. When there are five or more infection, the leucocytosis is brought young cells in the blood, ? I lobes connected by four or more fila- by appearing separate number and the of these ments, or there are five or more completely proportion depend!? the nature of the stimulus and the reacts? separate lobes. upon power of the marrow of the individual, and the The count is made and & by identifying classify- is a shift to the count. left in the Arneth _ .g one hundred neutrophil granulocytes from ing leucocytosis as a result of muscular exercise of the and the result is I different parts slide, only due to redistribution of is not accoi1 form :? cells, in the following . usually reported panied by any increase of young leucocytes 11 Example 1, a normal count the peripheral circulation, and is therefore associated with ' Class I II III IV V Total any left shift'. e in chronic infection or the Number .. 10 25 47 16 2 100 Again, toxaemia, is an increased leucocyte destruction, which nw there is an increase in the number of When or may not be balanced by new leucocyte form9 cells with one and two separate nuclear lobes, tion; in the former case there will be leucocyte? i.e., of class I or class II, at the expense of those or a normal count and in the latter leucopen1 with three or more lobes, there is said to be a In either case there is an increase in 1 y?Uw| left shift' in the Arnetli count, as in the forms, actual or relative, which results in a sn1 example given below :? to the left and a decrease in the weighted me? ^ It will be that this shift is Example 2, a 'left shift' count apparent quite ^ dependent of the total leucocyte or total gra1111 Class I II III IV V Total ' ' 0 locyte count, and provides another angle Number .. 41 44 14 1 0 100 the degree of the toxaemia. Conversely, when there is an increase in the In pernicious anaemia and certain other con^is 4- and multi-lobed nuclei, it is known as a tions of bone marrow dysfunction, there ^ ' right shift \ shift to the right in the Arneth count, &1! therefore an increase in the mean; ' weighted a count ^ Example 3, right shift' such conditions there are many multi-lobulate Class I II III IV V Total Normals Number 9 22 36 23 10 100 11 III IV Weight Various indices and methods of expressing mefiD more concisely the results of the Arneth count (Cooke's modification) have been proposed. Cooke and Ponder Probably the most suitable of these methods is (1927). the calculation of the weighted mean of the Kennedy (1933) (Iraq) nuclear this is obtained lobes; by multiplying Das Gupta (Calcutta) 15 2 the number of cells in class I by 1, the number in class II by 2, the number in class III 3, by cells and only very few young cells coming the number in class IV the number in by 4, the circulation. 1941] HEMATOLOGICAL TECHNIQUE : NAPIER & DAS GUPTA 99 phases, neutrophilic struggle, monocytic defence, B. Schilling count and lymphocytic cure, but it does not seem to take into account the nature of the infection In this count the neutrophil granulocytes classified and the variations in the divided into four groups and consequent response. there are so many directions in which follows :? Further, changes may take place, that the hsemogram *den- seems to facile >.ave a distinct peculiarly susceptible interpreta- 2. to the conscious or unconscious (. correspo tion suit wishes tation in the nucleus; they of the interpreter. a C- or We do not propose to give a full discussion 3. these have Stab?celisS'(St); on the interpretation of the hsemogram, as it has but no true shoe-shaped nucleus lo^latl^'count, anaJ only an indirect bearing on the subject of to class I of the Arneth correspond anaemia, but, as far as the neutrophil granulocyte to our staff or cells. ?0r?tive elements are concerned, Schilling's contention is Stab cells be of the rege may that infection may cause two types of reaction, and are differentiated ^ ^generative type, of the ^^ the regenerative reaction and degenerative re- and staining reaction in the new leuco- an cn'0tic,, action; regenerative reaction, ^hapethe latter the nucleus is ribbon-like py . are formed in the bone marrow and this is With and usua cytes clumps of chromatin, y^ reflected in the circulation a rise may be vacuola peripheral by shaped, and the cytoplasm in the total white-cell count, an increase in the Arneth.Arneth. juveniles and regenerative forms of stab cells, M III IV while in the degenerative reaction, which is found in severe toxaemia, on account of the degenerative influence of the toxin on the marrow, the formation of new cells is retarded; this may be shown by a leucopenia. In the differential count of such a case there are few juvenile cells, and the stab cells that reach the circulation fail to segment at the usual rate, and may degenerate with the result that there is a preponderance of stab forms, including many degenerative forms, in the blood. The reaction, (Reg.) however, may not be purely regenerative or degenerative in character, but may be a mixture of the two, and the hsemogram may be inter- mediate, with slight leucopenia and a greater (Dej) preponderance of the younger cells than in a J St.
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