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282 THE BLOOD.
by the aid of special appliances (L. Lewin, Miethe, and Stenger) the
band first described by Soret and then by Gamgee in the ultra-violet
portion. This violet band, X = 415, is of importance in the detection
of very small quantities of blood. While the two oxyhemoglobin bands
are still detectable in a dilution of 1 : 14, 700 the violet band may be seen,
according to Lewin, Miethe and Stenger l
in a dilution of 1 :40,000.
The observation of Piettre and Vila that so-called laky blood and oxyhsemo-
globin solutions in thick layers also show a third band in the red (X = 634) depends
in all probability, as also claimed fey Ville and Derrien, upon a partial forma-
tion of niethsemoglobin winch according to Aron 2
exists preformed in all blood.
A great many methods have been proposed for the preparation of
oxyhemoglobin crystals, but in their chief features they all agree with
the following one suggested by Hoppe-Seyler: The washed blood-
corpuscles (best those from the dog or the horse) are stirred with 2
vols, of water and then shaken with ether. After decanting the ether and
allowing the ether which is retained by the blood solution to evaporate
in an open dish in the air, cool the filtered blood solution to 0° C, add
while stirring one-fourth vol. of alcohol also cooled, and allow to stand
a few days at —5° to —10° C. The crystals which separate may be
repeatedly recrystallized by dissolving in water of about 35° C, cooling,
and adding cooled alcohol as above. Lastly, they are washed with
cooled water containing alcohol (one-quarter vol. alcohol) and dried
in vacuum at 0° C. or a lower temperature.3
For the preparation of oxyhemoglobin crystals in small quantities
from easily crystallizable blood, it is often sufficient to stir a drop of
blood with a little water on a microscope slide and allow the mixture
to evaporate so that the drop is surrounded by a dried ring. After
covering with a cover-glass, the crystals gradually appear radiating from
the ring. These crystals are formed more surely if the blood is first
mixed with some water in a test-tube and shaken with ether, and a drop
of the lower deep-colored liquid treated as above on the slide.
Haemoglobin, also called reduced haemoglobin or purple cruorin
(Stokes 4
), occurs only in very small quantities in arterial blood, in
larger quantities in venous blood, and is almost the only blood-coloring
matter after asphyxiation.
Hemoglobin is much more soluble than the oxyhemoglobin, and
it can therefore be obtained as crystals only with difficulty. These
1
Soret, cited in Maly’s Jaresb., 8; Gamgee, Zeitschr. f. Biol., 34; Lewin, Miethe
and Stenger, Pfliiger’s Arch., 118; Lewin and Miethe, ibid., 121.
2
Piettre and Vila, Compt. Rend., 140; Ville and Derrien, ibid., 140; Aron, Biochem.
Zeitschr., 3.
3
In regard to the preparation of oxyhemoglobin, see also Hoppe-Seyler-Thier-
felder’s Handbuch, 8. Aufl.; also the works cited in footnote 1, p. 278; also Schuur-
manns-Stekhoven, Zeitschr. f. physiol. Chem., 33, 296; see also Bohr, Skand. Arch,
f. Physiol., 3; J. Offringa, Bioch. Zeitschr., 28.
* Philosophical Magazine, 28, No. 190, Nov., 1864.
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