There are still therefore many contradictions in these results, and it is evidently necessary in a scientific investigation always to give the specific gravity of the serum and of the corpuscles, in addition to that of the total blood.

A method closely related to the estimation of the specific gravity is the direct estimation of the dried substance of the total blood, "Hygræmometry"; the clinical introduction of which we owe to Stintzing and Gumprecht. This method is really supplementary to those so far mentioned, and like them can be carried out with the small amounts of blood obtainable at the bedside without difficulty. Small quantities of blood are received in weighed glass vessels: which are then weighed, dried at 65°-70° C. for 24 hours and then weighed again. The figures so obtained for the dried substance have a certain independent importance; for they do not run quite parallel with those of the specific gravity, amount of hæmoglobin or number of corpuscles. The normal values are, for men 21.26%, for women 19.8%.

A further procedure for obtaining indirect evidence of the amount of hæmoglobin is the determination of the volume of blood corpuscles in 100 parts of total blood. For this estimation a method is desirable, which allows of the separation of the corpuscles from plasma in blood, that is as far as possible unaltered. The older methods do not fulfil this requirement; since they recommend either defibrination of the blood (quite impossible with the quantities of blood which are generally clinically available); or keeping it fluid by the addition of sodium oxalate or other substances which prevent coagulation. The separation of the two constituents can be effected by simply allowing the blood to settle, or with the centrifugal machine, specially constructed for the blood by Blix-Hedin and Gärtner ("Hæmatocrit").

For these methods various diluting fluids are used, such as physiological saline solution, 2.5% of potassium bichromate and many others. According to H. Kœppe they are not indifferent as far as the volume of the red blood corpuscles is concerned; and a solution which does not affect the cells must be previously ascertained for each specimen of blood. For this reason attention may be called to the proceeding of M. Herz, in which the clotting of the blood in the pipette is prevented by rendering the walls absolutely smooth by the application of cod-liver oil. Kœppe has slightly varied this method; he fills his handily constructed pipette, very carefully cleaned, with cedar wood oil, and sucks up the blood, as it comes from the fingerprick into the filled pipette. The blood displaces the oil, and as it only comes into contact with perfectly smooth surfaces, it remains fluid. By means of a centrifugal machine, of which he has constructed a very convenient variation, the oil as the lighter body is completely removed from the blood; and the plasma is also separated from the corpuscles. Three sharply defined layers are then visible, the layer of oil above, the plasma layer, and the layer of the red blood corpuscles. In as much as the apparatus is calibrated, the relation between the volumes of the plasma and corpuscles can be read off. No microscopical alterations in the corpuscles are to be observed.

Though this procedure seems very difficult of execution, it is nevertheless the only one, which has really advanced clinical pathology. The results of Kœppe—not as yet very numerous—give the total volume of the red corpuscles as 51.1-54.8%, an average of 52.6%.

M. and L. Bleibtreu have endeavoured indirectly to ascertain the relation of the volume of the corpuscles to that of the plasma. Mixtures of blood with physiological saline solution in various proportions are made, in each the amount of nitrogen in the fluid which is left after the corpuscles have settled is estimated. With the aid of quantities so obtained they calculate mathematically the volume of the serum and corpuscles respectively. Apart from the fact that a dilution with salt solution is also here involved, this method is too complicated and requires amounts of blood too large for clinical purposes. Th. Pfeiffer has tried to introduce it clinically in suitable cases, but has not so far succeeded in obtaining definite results. That, however, the relations between the relative volume of the red corpuscles and quantity of hæmoglobin are by no means constant, is well shewn by conditions (for example the acute anæmias) in which an "acute swelling" of the individual red discs occurs (M. Herz), but without a corresponding increase in hæmoglobin. The same conclusion results from recent observations of v. Limbeck, that in catarrhal jaundice a considerable increase of volume of the red blood corpuscles comes to pass under the influence of the salts of the bile acids.

As we have several times insisted, the quantity of hæmoglobin affords the most important measure of the severity of an anæmic condition. Those methods which neither directly nor indirectly give an indication of the amount of hæmoglobin are only so far of interest that they possibly afford an elucidation of the special pathogenesis of blood diseases in particular cases. To these belong the estimation of the alkalinity of the blood, which in spite of extended observations has not yet obtained importance in the pathology of the blood.

A value to which perhaps attention will be more directed than it has up to the present time by clinicians is the rate of coagulation of the blood, for which comparative results may be obtained by Wright's handy apparatus, the "Coagulometer." In certain conditions, particularly in acute exanthemata, and in the various forms of the hæmorrhagic diathesis, the clotting time is distinctly increased, or indeed clotting may remain in abeyance. Occasionally a distinct acceleration in the clotting, compared with the normal, may be observed. Wright has further ascertained in his excellent researches, that the clotting time can be influenced by drugs: calcium chloride, carbonic acid raise, citric acid, alcohol and increased respiration diminish the clotting power of the blood.

Recently Hayem has repeatedly called attention to a condition, that is probably closely connected with the coagulability of the blood. Although coagulation has set in, the separation of the serum from the clot occurs only very slightly or not at all. Hayem asserts, that he has found such blood in Purpura hæmorrhagica, Anæmia perniciosa protopathica, malarial cachexia: and some infectious diseases.

For such observations large amounts of blood are needed, which are clinically not frequently available. Certain precautions must be observed, as has been ascertained in the preparation of diphtheria serum, so that the yield of serum may be the largest possible. Amongst these that the blood should be received in longish vessels, which must be especially carefully cleaned, and free from all traces of fat. If the blood-clot does not spontaneously retract it must be freed from the side of the glass with a flat instrument like a paper-knife without injuring it. If no clot occurs in the cold, a result may perhaps follow at blood temperature.