US2673789A - Applicator for hemolyzing whole blood - Google Patents

Description

March 30, 1954 3 BROWN APPLICATOR FOR HEMOLYZING WHOLE BLOOD Original Filed Nov. 10, 1945 INVENTOR MO/PDEN G. ROWN TbRNsYs Patented Mar. 30, 1954 APPLICATOR FOR HEMOLYZING' WHOLE BLOOD Moi-den G. Brown, Darien', Conn a'ssignor to American Optical Company,

Soutlibridge,

Mass a voluntary association oi Massachusetts Original application November 10, 1945, Serial No. 627,873. Divided and this application August 1, 1950, Serial N0. 177,094

1 Claim.

This application is a division of copending appliea ticn Serial No. 627,873, which was filed No f fer 10, 19 -net; U. s. Patent No. 2,519,997, iss'lied August 22, 1950, and which relates to a. method for conveniently, precisely and rapidly determining the hemoglobin concentration of blood. The present application relates to apparams for use therewith.

Various conditions of a patients blood have a direct and material influence upon the health of the patient. Accordingly, accurate measurements of any of these conditions of the blood are highly desirable as, for example, measurements or the hemoglobin concentration in the blood which may be used to accurately indicate a normal condition or an abnormal condition of the patients blood such as anemia.

Since the hemoglobin concentration or red pigmerit of the blood is directly associated with the patients Ieoiiditioi'i, measurements of said con- A eentration are widely used by physicians for diag' noses and for accurate records of individual case histories. Such determination of the concentrau e f hemoglobin in the blood is generally made colorimetric'ally and cannot be made as aocurateiv with untreated whole blood as with hemslyzed blood since the variations in the number of blood cells, size and shape have a direct bearing there on. A colorimetric determination of the hemo= globin concentration of the" blood, accordingly, may be best had when the res pigment thereof is evenly distributed throughout the blood specimen and not conoentrated in cells separated by clear plas'ma. Accordingly, it is an object of this invention to provide improved means for subjecting a speci men of blood, which is in its most convenient form, oiryhemog'lobin, produced merely by exposure to air, to a treatment to hemolyze the said specimen to bring about a uniform distribution of red pigment or hemoglobin therein. It is a further object of the invention to provide means to foifn this specimen into oxyhern'oglobin which may be formed as a layer of uniform predeter= inined thickness whereby the light-absorbing characteristics of this specimen may be studied and compared against a comparison member or a series of comparison members having known light-absorbing values, to thereby obtain ah accurate indication of the hemoglobin condition oi the patients blood. I

It is also an object of the invention to provide convenient means in the form ofa rod or stick of suitable material having a handle portion and diapered end portion for receiving a coating of 2 an hemolytic agent thereon and for supporting said coating while being applied to the blood specimen being hemolyzed.

Another object of the invention is to provide such a rod or stick coated with a blood hemolyzing agent on a tapered portion thereof was to be supported by the stick in spaced relation to a supporting surface, or the like, and to be spaced from other-similar stick when bunched together in a container for preventing adhesion between the coatings thereon prior to their use.

Further objects and advantages of the invention will become apparent from a consideration of the detailed description which follows taken in conjunction with the accompanying drawings wherein a pre'ferredform of the apparatus and certain steps in the method to be'emplcyed therewith are shown and described. It will be understood, however, that the invention is not to be limited to the details of the disclosure except as defined by the appended claim since the disclosure is given for purposes of illustration only.

In the drawings:

Fig. 1 is a perspective view of a pair of plates which may be used with the apparatus of the present invention, said plates being shown in spaced relation in order to disclose the details of construction thereof;

Fig. 2 is a perspective view of the plates of Fig. 1 shown in clamped relation in a resilient holder and ready for use;

Fig. 3 is a perspective View of the plates likewise positioned in the holder, but having a specimen-receiving area of the lower plate offset or moved outwardly to a position for receiving-a specimen thereon, and with 2. treated stick or rod in position for stirring the specimen on the specimen-receiving plate;

Fig. 4 shows an enlarged view of the treated stick of the invention; I

Fig. 5 shows a carrying case or container for receiving a plurality of said sticks or rods, said container forming a substantially moisture proof enclosure therefor; and

Fig. 6 is an enlarged fragmentary view showing two similar specimen-receiving plates placed to-' gether for use in forming a blood chamber of double thickness.

Referring to the drawings in detail, and par ticularly Fig. 1, the numeral ill indicates generally a lower specimen-receiving plate which may be used in'combination with a cover plate M of equal size, general shape and thickness. Both of said plates are transparent. The upper and lower surfaces i4 and it of the upper plate are accurately finished to provide optically flat surfaces or parallel planes thereon. The lower plate In is likewise finished upon it lower surface l8 and has its upper laterally spaced side rail portions 29 and 22 accurately formed so as to be parallel to and equally spaced from the bottom surface l8. The surfaces 20 and 22 are laterally spaced from a pair of intermediate specimenreceiving areas 2 3 and 25 by means of grooves 26. and said area are also spaced from each other by an intermediate groove 28. The grooves 26 and 28 are of a depth sufficient to form spaces for receiving excess blood from the area 24 or 25 when the plates 10 and I2 are assembled one upon the other. The areas 24 and 25 for receiving the blood specimen til, as indicated in Fig. 3, are accurately formed so as to be disposed a predetermined distance below the plane of the side rail portions 20 and 22, and are arranged parallel to said plane, thereby forming a space of uniform predetermined thickness between each of these areas and the cover plate l2 when the cover plate is positioned thereover.

The plates l and I2 may be conveniently and firmly held in an assembled position by a clamp or holder 32 which is formed of resilient material, such as bronze, stainless steel or the like, and has a base portion 34 to which a handle 36 of suitable size and shape may be riveted or otherwise secured. The holder is provided with a pair of spaced resilient side members 38 extending in substantially parallel relation from the base portion 34, and one of said side members is provided with an upper pair of arms 40 and the other of said side members is provided with a lower pair of arms 40' arranged in laterally spaced relation to provide upper and lower unobstructed areas or spaces 42 aligned with specimen-receiving areas 24 and 25 through which the specimen 38 may be observed. The distance between the members 38 is somewhat greater than the combined thickness of the plates Ill and [2, as is clearly indicated in Fig. 2, so that the lower pair of arms 40' may be provided with inwardly curved end portions 44, as shown, to engage the outer surface of one of the plates and press both plates into firm engagement with the other pair of resilient arms 40. Thus, when these plates are in an assembled position in the holder, they will be firmly clamped together and provide between the area 24 or 25 and the cover plate a space of predetermined thickness. A pair of indentations 65 are pressed into the base portion 34 near opposite ends thereof so that they will be engaged by the inner edges of the plates GE! and [2 when in the holder 32 and space these plates from the inner wall or surface of the base portion 34 thus preventing any blood which may be on these edges from touching and adhering to said inner surface.

Fig. 3 shows the specimen plate and cover plate 12 in an assembled position clamped between the resilient arms 46, 40' of the holder 32 and the specimen plate it is positioned so that the specimen-receiving area 24 is exposed and may have placed thereon a blood specimen 3!). A treated rod or stick 45 is shown in contact with the area 24 and the specimen 36 thereon for a purpose to be presently described. The rod or stick at is best shown in Fig. 4 and may be formed of a suitable stiff or rigid inert material, such as wood. It comprises an elongated handle portion 48 provided with straight parallel sides which extend the greater portion of the length of the stick and a tapered portion 50 adjacent an end 4 thereof as indicated at 5| so as to provide a reduced end portion for receiving an hemolytic agent 52. By the words inert material as used above and hereinafter in the specification and claims is meant a material which will not be materially affected by the hemolytic agent forming the coating 52 and which will not alter the condition of the blood specimen under examination.

The rod or stick may be made of any preferred cross-sectional shape desired, but as shown is circular. While the proportions or dimensions of this rod or stick may vary somewhat, a preferred size would be between 1% and 1% inches in length and from to of an inch in thickness. The tapered end portions 50 of the rod or stick 46 are formed so as to receive the coating 52 upon the outer end thereof and support said coating in a space entirely within the planes containing the parallel sides of the stick, whereby the hemolytic material will be normall spaced from or above any supporting surface upon which the stick rests, or spaced from the side Walls of a container or the like used therewith, or spaced from other similar sticks when bunched together in said container.

A suitable hemolytic agent which has been found to give satisfactory result for forming the layer or coating 52 may comprise saponin which may be applied to the stick by forming a tacky solution of saponin in water or other suitable solvent and dipping the ends of the rods or sticks 46 into said solution. The coating on the rod or stick is then air dried or dried in an oven at temperatures not exceeding 200 F. Improved results may be obtained by adding a small amount of sodium oxalate or other anti-coagulant to the hemolytic agent to be applied to the rods or sticks, which additional material will materially lessen the tendency of the blood specimen to coagulate.

It is desirous that the treated rod or stick 46 be formed of stiff, durable and inert material in order that the material forming the hemolytic agent will not be affected thereby. A convenient amount of hemolytic agent upon the tapered end of the stick has been found to be provided by dipping each stick to a depth of from to A of an inch into said hemolytic solution with or without the anti-coagulant as desired. A container 54 is shown in Fig. 5, said container being formed of a substantially moisture proof plastic material to serve as a suitable means for storing and transporting a plurality of sticks as part of the equipment of the invention. Relative long tapered portions are formed at 56 and 58 on the two parts of this separable container in order to provide a substantially air tight joint between the two parts, in order that moisture or dampness of the outside air may be excluded from the container and thus not affect the condition of the hemolytic material upon the ends of the sticks contained therein.

A preferred method of use of the apparatus described above for the determination of the hemoglobin content or concentration of the blood specimen comprises puncturing the finger or lobe of the ear of the patient to obtain a small quantity of blood to be tested, said blood being placed upon the area 24, or 25, of the plate I0, when in a separated condition as shown by Fig. 1 or when in an assembled position with the plate [0 offset or projecting outwardly of the plate l2 and gripped by the resilient arms of the holder as shown by Fig. 3, and this may be done by merely touching the area 24, or 25, to the finger or ear lobe. The specimen 30 so obtained on the specimen-receiving area is then agitated or stirred by the treated end of the stick 46. When the specimen is subjected to the hemolyzing agent, such as sap-onin, for a sufficient time during the stirring operation, such as for a period ranging from five to fifteen seconds, a hemolyzing action upon the blood specimen will be produced, which action brings about a rupturing of the membranes surrounding the individual red cells or corpuscles of the blood, allowing the red pigment or hemoglobin thereof to be evenly distributed throughout the specimen and thereby producing a solution of substantially uniform hemoglobin concentration.

The plate iii containing the hemolyzed solution so produced may then be covered by the cover plat 12 and pressed or slipped into place in the holder 32, as shown by Fig. 2, or may, if in the oiTset position shown in Fig. 3, be pressed inwardly to a position adjacent the cover plate i2 so that, in either case, the prepared solution will be contained between the lower surface Id of the plate E2 and the area 24, or 25, of the plate it as a uniform layer of predetermined thickness. The solution so prepared will tend to spread over the entire specimen-receiving area by capillary attraction between these adjacent slightly spaced surfaces. Any excess solution at the outer edge of the plates Ill and I2 may be wiped therefrom and if there is an excess amount on the area when the cover plate is placed over the area 24, or 25, it may flow into the grooves 25 or 28 or to the outer edges of the plates [0 and 12 as the layer of predetermined thickness is formed.

When a layer of uniform thickness of prepared solution has been produced between the plates 12 and 52, these plates may be positioned in the field of a colorimetric device or the like and the light-absorption properties thereof observed and compared with a standard comparison member or a series of standard comparison areas of known light-absorbing values. Thus when a comparison area of known value and of lightabsorbing properties which are the same as the specimen has been ascertained an accurate indication of the hemoglobin concentration of the specimen may be determined therefrom.

In cases of extreme anemia wher the amount of red pigment or hemoglobin in the blood is relatively small the physician may employ to good advantage two specimen-receiving plates ill with the specimen-receiving area thereon facing each other as shown by Fig. 6, thereby forming blood chambers between each adjacent pair of areas of double predetermined thickness as indi cated by the letters t and t and, accordingly, the specimen placed therebetween will have double light-absorbing values which may be, in said cases of extreme anemia, more accurately observed and measured or compared in a colorimetric device, or the like, than could the specimen if it were only half as thick. Obviously, in such a combination of similar plates a blood chamber of single thickness t may readily b formed when 6 desired by merely placing the upper plate upside-down upon the lower plate since surfaces 2 1 and 25 are parallel to and equally spaced from the surface).

Thus, it will be seen that a convenient, accurate and enici-entmethod and apparatus have been shown and described for ascertaining the hemoglobin content or concentration of a patients blood.

Having described the invention, I claim:

A preformed applicator for use in hemolyzing a minute specimen of freshly extracted whole blood which is to be tested for determining the hemoglobin concentration thereof, said applicator comprising an elongated rod of rigid and durable material provided with a handle portion having approximately straight parallel sides which extend the greater portion of the length of the rod and terminate in a relatively short portion tapered toward the end thereof and comprising the remainder of said rod, said tapered portion being coated with a thin dried adherent layer of hemolytic agent comprising saponin which extends from said end of the rod partially along the length of the tapered portion and being sufiiciently thin that the hemolytic agent lies substantially entirely within the planes containing the parallel sides of the rod whereby a plurality of such applicators may be grouped with their handle portions in contiguous side-by-side relation with each other while maintaining the hemolytic agent on said tapered portions spaced from each other, said tapered portion, at least, of the rod being formed of a material which will not be materially affected by and which will not materially affect the hemolytic agent.

MORDEN G. BROWN.

References Cited in the file of this patent UNITED STATES PATENTS Number Name Date 174,619 Clark Mar. 14, 1876 410,794 Hellwig Sept. 10, 1889 458,244 Tyree Aug. 25, 1891 1,498,073 Cohn June 1'7, 1924 1,969,874 Butterfield Aug. 14, 1934 2,008,206 Grant July 16, 1935 2,206,282 Jacobius July 2, 1940 FOREIGN PATENTS Number Country Date 496,345 Great Britain Nov. 29, 1938 OTHER REFERENCES Wintrobe, Clinical Hematology, 2nd ed.,

1946, pages 127, 241, 274, 275. Lea and Febiger, Philadelphia, publishers.

Suffer, L. J., Medicine, vol. 14, 1935, page 185.

Heller et al., Jour. Lab. and Clin. Med, vol. 19, 1934, page 777.

.Todd et al., Clinical Diagnosis by Laboratory Methods, 9th ed., 1939, pages 248-251, 320, 321, 347; W. B. Saunders 00., Philadelphia, publishers.

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