- Email: [email protected]
An apparatus for the intracavity administration of radioactive colloidal gold
266
Technical notes
(A) t h r o u g h a 10 m m glass t u b i n g connection (B) into the b o t t o m of the cage. T h e air left the cage t h r o u g h a 10 m m glass t u b i n g connection c o n t a i n e d in a r u b b e r stopper in a n opening n e a r the top of the cage (E). T h e air t h e n passed to a series of tubes for the collection of C O 2. Pyrex test tubes, 300 × 35 m m , fitted w i t h r u b b e r stoppers into w h i c h were placed inlets a n d outlets from glass manifolds, were used for the collection of C O 2. T h e inlet tube (F) was p r o v i d e d w i t h a stopcock above, the r u b b e r stopper. Below the stopper, it was connected w i t h tygon t u b i n g to a gas dispersion tube ( m e d i u m porosity, 20 ram) w h i c h e x t e n d e d below the surface of 0' 1 N sodium hydroxide. T h e dispersion disc was suspended in a vertical plane. A second hole in the r u b b e r stopper cf the collection tube enclosed a Pyrex t u b i n g a r m w h i c h led to a second tube in the set, e q u i p p e d in the same m a n n e r as the first. T h e outlet of the second tube (G), however, was connected b y means of a stopcock to the second manifold. E a c h manifold c o n t a i n e d connections to four such sets of tubes. This p e r m i t t e d collection of C O z in the two tubes of each set i n d e p e n d e n t l y of the other tubes a n d m a d e possible the i n t e r m i t t e n t collection of samples d u r i n g a n y desired time interval w i t h o u t loss of C O 2. T h e second manifold was connected with tygon t u b i n g to a t r a p (H) a n d thence to a n aspirator to provide a source of suction. I n our work it was desirable to connect four sets of two tubes to the manifolds because of the short time intervals for the collection of C O 2 d u r i n g the early p a r t of the experiments. However, one set of two tubes will suffice in instances where one total collection is desired. T h e cost of all materials used in the construction of the metabolism apparatus, using only one set of two C O 2 collection tubes, did not exceed $20, whereas commercially available m e t a b o l i s m cages of similar construction for rats cost a p p r o x i m a t e l y $300. T h e parts of the apparatus, with the exception of the Plexiglas, are n o r m a l l y found in chemistry laboratories. T h e hole in the r u b b e r stopper (B) c o n t a i n i n g the tail Of the r a t c a n be plugged w h e n it is not necessary to collect blood samples. T h e a p p a r a t u s c a n be checked for air-tightness prior to use by placing a small beaker c o n t a i n i n g a few ml of c o n c e n t r a t e d hydrochloric acid on the floor of the cage a n d passing a beaker of a m m o n i u m hydroxide over the junctions, as gentle suction is applied. I f a cloud of a m m o n i u m chloride v a p o r forms at a n y Ple×iglas j u n c t i o n , it c a n be resealed with the Plexiglas cement. T h e authors found the cage to be perfectly airtight w h e n the lid was sealed with adhesive tape.
T h e rate of flow of air is regulated b y the flow of water t h r o u g h the aspirator. T h e cage was o p e r a t e d in a n air-conditioned room. W h e r e this convenience is not available, a t h e r m o m e t e r c a n b e inserted t h r o u g h one of the r u b b e r stoppers a n d the t e m p e r a ture controlled w i t h fans a n d moist towels in difficult w e a t h e r conditions. Additional holes c a n be drilled as desired for other purposes. T h e cage floor a n d all other components c a n be separated for easy cleaning. This investigation was s u p p o r t e d in p a r t b y a research g r a n t from the N a t i o n a l Institute of Arthritis a n d M e t a b o l i c Diseases, N a t i o n a l Institutes of Health, Public H e a l t h Service. T h e a n i m a l experiments will be p u b l i s h e d at a later date. GERALD A. EDWARDS
Department of Chemistry CECILE H. EDWARDS Department of Home Economics EVELYN L. GADSDEN The Agricultural and Technical College of North Carolina, Greensboro, North Carolina
Reference l. ROTH L . J .
Nucleonics 14, 104 (1956).
An A p p a r a t u s for the I n t r a c a v i t y Administration of Radioactive Colloidal Gold (Received 27 March, 1958; in revisedform 1 September, 1958) AN earlier p a p e r (1) described e q u i p m e n t for the intracavity a d m i n i s t r a t i o n of radioactive colloidal gold. W i t h this e q u i p m e n t it was possible to dilute a sterile p r e p a r a t i o n of gold aseptically to a n y volume a n d to r u n it into the p a t i e n t b y gravity feed, the whole operation b e i n g remotely controlled. Experience with this e q u i p m e n t suggested certain i m p r o v e m e n t s w h i c h m i g h t be m a d e o n grounds of r a d i a t i o n protection. These relate principally to the m e t h o d of r e m o v i n g the gold from the a m p o u l e in w h i c h it is delivered a n d to the taps used to control the flow of radioactive solution. These i m p r o v e ments, a n d other m i n o r ones, h a v e now been incorp o r a t e d in the a p p a r a t u s described here, the working of w h i c h m a y be understood by reference to Fig. 1. T h e essential glass p a r t is a reservoir,* of v o l u m e a b o u t 100 ml, w i t h a ground-glass L u e r nozzle a n d a side-arm. T o the side-arm is a t t a c h e d a length of silicone-rubber t u b i n g b e a r i n g a s p h y g m o m a n o m e t e r b u l b with_ its air-intake p o r t stopped. T h e lower outlet tube of the reservoir carries a T-piece, one * Specially made by Messrs. Aimer Ltd.
Technical notes
arm of which leads to the patient, whilst saline can be introduced into the reservoir to any degree required by means of the other. T h e reservoir, with its attendant tubes and T-piece is sterilized as a unit together with a Luer serum needle of suitable dimensions in situ on the nozzle. The two taps referred to in Fig. 1 as "quick-action" form one of the novel features of this apparatus. Their working may be understood by reference to Fig. 2. Each consists of a clamp A which can be
Salln~e B~~a
~H
d
H
K
MI 1' 22L
bulb
~ TopatTent
Fro. l. General arrangement.
I r. . . . . . . . . .
;
,I Jl
il I',
O
267
opened or closed by action of a cam, (shaded) which is itself actuated by means of half a turn of the operating handle. The clamp m a y be held firmly in any position (fully open or shut, or at any point intermediate between the two) by means of the friction provided by a cork friction-plate B loaded against the inside face of the case by means of a spring C. A forked slide (not shown in diagrams) holds the rubber tube in the correct position under the clamp. T h e second major improvement concerns the mechanism by which the gold solution is extracted from the sterile ampoule in which it is delivered. This m a y be understood by reference to Figure 1 and the accompanying photograph. T h e ampoule is held upside-down in forceps and placed in the Perspex carrier A where it is firmly held by the bayonet cap a, which carries a sponge rubber pad B fm the purpose. T h e carrier is brought down onto the needle point by half a turn of the operating handle (shown in the photograph) ; this is also sufficient to cause the needle to penetrate the rubber ampoule-cap. T h e m o v e m e n t is transmitted from the operating handle by means of the cam D moving in the rocker-arm about the pivot E ; the degree of m o v e m e n t required can be adjusted by means of the movable stops F. T h e Perspex carrier is pivoted at G in the yoke H and supported vertically in the bearing I. When the ampoule-carrier is at its highest point its m o v e m e n t is held by a spring-loaded ball K in a dimple on the operating cam. This mechanism is very robustly built since the rubber caps supplied on U . K . A . E . A . ampoules are much harder to puncture than those on normal pharmaceutical ampoules, a fact which can be a source of hazard unless special precautions are taken. T h e needle itself is locked into a metal clamp J . W h e n the needle has punctured the rubber cap, the reservoir being empty, the sphygmomanometer bulb is compressed. Air is forced up the needle and into the ampoule; and on releasing the bulb some of the radioactive solution is sucked into the reservoir. Repetition of this operation 3 to 6 times transfers all the gold aseptically to the reservoir. T h e
FIa. 2. Quick-action tap.
Technical notes
268
air-release valve o n the s p h y g m o m a n o m e t e r b u l b is t h e n unscrewed to act as a n airway, after w h i c h saline c a n be r u n in to dilute the colloidal gold to the e x t e n t required. I t m a y be readily observed from the colour of the gold solution t h a t mixing is instantaneous a n d complete. T h e diluted gold solution is then r u n into the p a t i e n t a t w h a t e v e r speed is required together w i t h such f u r t h e r washings of saline as m a y be necessary. T h e a p p a r a t u s is m o u n t e d on a b o a r d the lower edge of w h i c h is at a height of a p p r o x i m a t e l y 5 it, the saline bottle b e i n g at a b o u t 6 ft 6 in. ; this gives sufficient h e a d of pressure for efficient r u n n i n g cf the p r e p a r a t i o n into the patient. T h e e q u i p m e n t is fully shielded b y lead when in operation, a n d observation a n d control of the process are m a i n t a i n e d b y indirect vision using a l a m p house a n d mirror. I t is claimed t h a t incidental r a d i a t i o n h a z a r d to the o p e r a t o r has b e e n eliminated as fully as practicable in this e q u i p m e n t .
Reference 1. ARNOTT D. G. a n d JoY A. R. 29, 337 (1956).
The London Hospital London E1
Brit. J. Radiol. D . G . ARNOTT H . R . KNIGHT
Effects of Ionizing Radiations
on Tissue
Mast Cells* (Received 16 May 1958) IT is believed t h a t the r a d i a t i o n sickness is due to liberation of histamine in the body. ~1~ H i s t a m i n e liberation is t h o u g h t to b e one of the m a i n functions of tissue mast cells. W i t h this idea in view, a n a t t e m p t was m a d e to see the effects of different kinds of radiations b o t h from external a n d internal sources on tissue mast cells. Results of this study form the subject m a t t e r of this c o m m u n i c a t i o n .
M e t h o d s and m a t e r i a l s T h e mesenteric spread of 40 a d u l t h e a l t h y albino rats weighing from 175 to 225 g were used in this study. T h e a b d o m e n was opened after killing the a n i m a l w i t h n e m b u t a l a n d a loop of j e j u n u m a n d * This paper was presented at the 7th Annual Conference of the Indian Association of Pathologists held in November 1956 at Mysore (South India).
ileum was d r a p e d over a slide. O n c e the mesentery a n d its vessels h a d a d h e r e d to the glass the guts a n d the mesenteric a t t a c h m e n t s were cut away. All the slides were fixed in absolute alcohol for 6 h r a n d stained with 0-25% toluidin blue in 35°o alcohol for 2-3 rain. O n each slide from each a n i m a l 20 fields, 4-4196 m m 2 each, were covered using a magnification factor of 150. T h e variation in the total mast cell c o u n t per u n i t area of mesenteric window along with the percentage of morphological variations were noted. M o r p h d o g i c a l characters were recorded with the camera lucida d r a w i n g a n d actual changes in the size were m e a s u r e d with a planimeter. A u t o r a d i o g r a p h i c studies of the peritoneal spread were done. Doses of isotopes were injected intraperitoneally. X-rays were given from 106 V a n d 250 kV machines.
Results Number. T a b l e 1 shows t h a t in the control g r o u p the average n u m b e r of mast cells per 20 fields was 310-4, This n u m b e r was increased in all the irradiated groups. R e m a r k a b l e increase was seen after irradiation with 1 0 6 V X-rays a n d Fe 59, n a m e l y 528 a n d 413 respectively. T h e average n u m b e r after t r e a t m e n t with ps2, Au 19s, a n d 250 kV X-rays did not show m u c h difference, the n u m b e r s being 344.7, 387, a n d 378.5 respectively. Size. T h e average area of cross section of the mast cells in control group was 4 2 3 . 8 / ~ . T h e rem a r k a b l e change was seen again for Fe ~9 with or without X-ray. Here most of the cells were very small, the average area being 151-1/~2 for Fe 59 a n d 140.3 #2 for Fe 59 along with 250 k V X-ray. T h e average area for p32 was 212.9/~ 2, for A u 19s 225.7 #2 a n d for b o t h 250 kV a n d 106 V X-rays 161-1 #2. Percentage of morphological variations. T h e various morphological characters observed were placed in 3 categories : 1. N o r m a l - - c e l l s with uniform distribution of m e t a c h r o m a t i c granules with or w i t h o u t visible nucleus. 2. Small h y p e r c h r o m a t i c - - c e l l s smaller t h a n the average of the above described n o r m a l cells a n d stained so deeply t h a t nuclei were not seen a n d the granules were found to be c l u m p e d together. 3. O t h e r s - - t h o s e which show g r a n u l a r shifting, bipolar giving a d u m b - b e l l shape, u n i p o l a r shifting of granules, small cells with nucleus filling almost the whole of the cells, cells with eccentric nucleus, cells with vacuolation a n d g r a n u l a r spilling, clumping of the granules in groups, cells h a v i n g peculiar shapes a n d cells showing degranulation. It is
Technical notes
(A) t h r o u g h a 10 m m glass t u b i n g connection (B) into the b o t t o m of the cage. T h e air left the cage t h r o u g h a 10 m m glass t u b i n g connection c o n t a i n e d in a r u b b e r stopper in a n opening n e a r the top of the cage (E). T h e air t h e n passed to a series of tubes for the collection of C O 2. Pyrex test tubes, 300 × 35 m m , fitted w i t h r u b b e r stoppers into w h i c h were placed inlets a n d outlets from glass manifolds, were used for the collection of C O 2. T h e inlet tube (F) was p r o v i d e d w i t h a stopcock above, the r u b b e r stopper. Below the stopper, it was connected w i t h tygon t u b i n g to a gas dispersion tube ( m e d i u m porosity, 20 ram) w h i c h e x t e n d e d below the surface of 0' 1 N sodium hydroxide. T h e dispersion disc was suspended in a vertical plane. A second hole in the r u b b e r stopper cf the collection tube enclosed a Pyrex t u b i n g a r m w h i c h led to a second tube in the set, e q u i p p e d in the same m a n n e r as the first. T h e outlet of the second tube (G), however, was connected b y means of a stopcock to the second manifold. E a c h manifold c o n t a i n e d connections to four such sets of tubes. This p e r m i t t e d collection of C O z in the two tubes of each set i n d e p e n d e n t l y of the other tubes a n d m a d e possible the i n t e r m i t t e n t collection of samples d u r i n g a n y desired time interval w i t h o u t loss of C O 2. T h e second manifold was connected with tygon t u b i n g to a t r a p (H) a n d thence to a n aspirator to provide a source of suction. I n our work it was desirable to connect four sets of two tubes to the manifolds because of the short time intervals for the collection of C O 2 d u r i n g the early p a r t of the experiments. However, one set of two tubes will suffice in instances where one total collection is desired. T h e cost of all materials used in the construction of the metabolism apparatus, using only one set of two C O 2 collection tubes, did not exceed $20, whereas commercially available m e t a b o l i s m cages of similar construction for rats cost a p p r o x i m a t e l y $300. T h e parts of the apparatus, with the exception of the Plexiglas, are n o r m a l l y found in chemistry laboratories. T h e hole in the r u b b e r stopper (B) c o n t a i n i n g the tail Of the r a t c a n be plugged w h e n it is not necessary to collect blood samples. T h e a p p a r a t u s c a n be checked for air-tightness prior to use by placing a small beaker c o n t a i n i n g a few ml of c o n c e n t r a t e d hydrochloric acid on the floor of the cage a n d passing a beaker of a m m o n i u m hydroxide over the junctions, as gentle suction is applied. I f a cloud of a m m o n i u m chloride v a p o r forms at a n y Ple×iglas j u n c t i o n , it c a n be resealed with the Plexiglas cement. T h e authors found the cage to be perfectly airtight w h e n the lid was sealed with adhesive tape.
T h e rate of flow of air is regulated b y the flow of water t h r o u g h the aspirator. T h e cage was o p e r a t e d in a n air-conditioned room. W h e r e this convenience is not available, a t h e r m o m e t e r c a n b e inserted t h r o u g h one of the r u b b e r stoppers a n d the t e m p e r a ture controlled w i t h fans a n d moist towels in difficult w e a t h e r conditions. Additional holes c a n be drilled as desired for other purposes. T h e cage floor a n d all other components c a n be separated for easy cleaning. This investigation was s u p p o r t e d in p a r t b y a research g r a n t from the N a t i o n a l Institute of Arthritis a n d M e t a b o l i c Diseases, N a t i o n a l Institutes of Health, Public H e a l t h Service. T h e a n i m a l experiments will be p u b l i s h e d at a later date. GERALD A. EDWARDS
Department of Chemistry CECILE H. EDWARDS Department of Home Economics EVELYN L. GADSDEN The Agricultural and Technical College of North Carolina, Greensboro, North Carolina
Reference l. ROTH L . J .
Nucleonics 14, 104 (1956).
An A p p a r a t u s for the I n t r a c a v i t y Administration of Radioactive Colloidal Gold (Received 27 March, 1958; in revisedform 1 September, 1958) AN earlier p a p e r (1) described e q u i p m e n t for the intracavity a d m i n i s t r a t i o n of radioactive colloidal gold. W i t h this e q u i p m e n t it was possible to dilute a sterile p r e p a r a t i o n of gold aseptically to a n y volume a n d to r u n it into the p a t i e n t b y gravity feed, the whole operation b e i n g remotely controlled. Experience with this e q u i p m e n t suggested certain i m p r o v e m e n t s w h i c h m i g h t be m a d e o n grounds of r a d i a t i o n protection. These relate principally to the m e t h o d of r e m o v i n g the gold from the a m p o u l e in w h i c h it is delivered a n d to the taps used to control the flow of radioactive solution. These i m p r o v e ments, a n d other m i n o r ones, h a v e now been incorp o r a t e d in the a p p a r a t u s described here, the working of w h i c h m a y be understood by reference to Fig. 1. T h e essential glass p a r t is a reservoir,* of v o l u m e a b o u t 100 ml, w i t h a ground-glass L u e r nozzle a n d a side-arm. T o the side-arm is a t t a c h e d a length of silicone-rubber t u b i n g b e a r i n g a s p h y g m o m a n o m e t e r b u l b with_ its air-intake p o r t stopped. T h e lower outlet tube of the reservoir carries a T-piece, one * Specially made by Messrs. Aimer Ltd.
Technical notes
arm of which leads to the patient, whilst saline can be introduced into the reservoir to any degree required by means of the other. T h e reservoir, with its attendant tubes and T-piece is sterilized as a unit together with a Luer serum needle of suitable dimensions in situ on the nozzle. The two taps referred to in Fig. 1 as "quick-action" form one of the novel features of this apparatus. Their working may be understood by reference to Fig. 2. Each consists of a clamp A which can be
Salln~e B~~a
~H
d
H
K
MI 1' 22L
bulb
~ TopatTent
Fro. l. General arrangement.
I r. . . . . . . . . .
;
,I Jl
il I',
O
267
opened or closed by action of a cam, (shaded) which is itself actuated by means of half a turn of the operating handle. The clamp m a y be held firmly in any position (fully open or shut, or at any point intermediate between the two) by means of the friction provided by a cork friction-plate B loaded against the inside face of the case by means of a spring C. A forked slide (not shown in diagrams) holds the rubber tube in the correct position under the clamp. T h e second major improvement concerns the mechanism by which the gold solution is extracted from the sterile ampoule in which it is delivered. This m a y be understood by reference to Figure 1 and the accompanying photograph. T h e ampoule is held upside-down in forceps and placed in the Perspex carrier A where it is firmly held by the bayonet cap a, which carries a sponge rubber pad B fm the purpose. T h e carrier is brought down onto the needle point by half a turn of the operating handle (shown in the photograph) ; this is also sufficient to cause the needle to penetrate the rubber ampoule-cap. T h e m o v e m e n t is transmitted from the operating handle by means of the cam D moving in the rocker-arm about the pivot E ; the degree of m o v e m e n t required can be adjusted by means of the movable stops F. T h e Perspex carrier is pivoted at G in the yoke H and supported vertically in the bearing I. When the ampoule-carrier is at its highest point its m o v e m e n t is held by a spring-loaded ball K in a dimple on the operating cam. This mechanism is very robustly built since the rubber caps supplied on U . K . A . E . A . ampoules are much harder to puncture than those on normal pharmaceutical ampoules, a fact which can be a source of hazard unless special precautions are taken. T h e needle itself is locked into a metal clamp J . W h e n the needle has punctured the rubber cap, the reservoir being empty, the sphygmomanometer bulb is compressed. Air is forced up the needle and into the ampoule; and on releasing the bulb some of the radioactive solution is sucked into the reservoir. Repetition of this operation 3 to 6 times transfers all the gold aseptically to the reservoir. T h e
FIa. 2. Quick-action tap.
Technical notes
268
air-release valve o n the s p h y g m o m a n o m e t e r b u l b is t h e n unscrewed to act as a n airway, after w h i c h saline c a n be r u n in to dilute the colloidal gold to the e x t e n t required. I t m a y be readily observed from the colour of the gold solution t h a t mixing is instantaneous a n d complete. T h e diluted gold solution is then r u n into the p a t i e n t a t w h a t e v e r speed is required together w i t h such f u r t h e r washings of saline as m a y be necessary. T h e a p p a r a t u s is m o u n t e d on a b o a r d the lower edge of w h i c h is at a height of a p p r o x i m a t e l y 5 it, the saline bottle b e i n g at a b o u t 6 ft 6 in. ; this gives sufficient h e a d of pressure for efficient r u n n i n g cf the p r e p a r a t i o n into the patient. T h e e q u i p m e n t is fully shielded b y lead when in operation, a n d observation a n d control of the process are m a i n t a i n e d b y indirect vision using a l a m p house a n d mirror. I t is claimed t h a t incidental r a d i a t i o n h a z a r d to the o p e r a t o r has b e e n eliminated as fully as practicable in this e q u i p m e n t .
Reference 1. ARNOTT D. G. a n d JoY A. R. 29, 337 (1956).
The London Hospital London E1
Brit. J. Radiol. D . G . ARNOTT H . R . KNIGHT
Effects of Ionizing Radiations
on Tissue
Mast Cells* (Received 16 May 1958) IT is believed t h a t the r a d i a t i o n sickness is due to liberation of histamine in the body. ~1~ H i s t a m i n e liberation is t h o u g h t to b e one of the m a i n functions of tissue mast cells. W i t h this idea in view, a n a t t e m p t was m a d e to see the effects of different kinds of radiations b o t h from external a n d internal sources on tissue mast cells. Results of this study form the subject m a t t e r of this c o m m u n i c a t i o n .
M e t h o d s and m a t e r i a l s T h e mesenteric spread of 40 a d u l t h e a l t h y albino rats weighing from 175 to 225 g were used in this study. T h e a b d o m e n was opened after killing the a n i m a l w i t h n e m b u t a l a n d a loop of j e j u n u m a n d * This paper was presented at the 7th Annual Conference of the Indian Association of Pathologists held in November 1956 at Mysore (South India).
ileum was d r a p e d over a slide. O n c e the mesentery a n d its vessels h a d a d h e r e d to the glass the guts a n d the mesenteric a t t a c h m e n t s were cut away. All the slides were fixed in absolute alcohol for 6 h r a n d stained with 0-25% toluidin blue in 35°o alcohol for 2-3 rain. O n each slide from each a n i m a l 20 fields, 4-4196 m m 2 each, were covered using a magnification factor of 150. T h e variation in the total mast cell c o u n t per u n i t area of mesenteric window along with the percentage of morphological variations were noted. M o r p h d o g i c a l characters were recorded with the camera lucida d r a w i n g a n d actual changes in the size were m e a s u r e d with a planimeter. A u t o r a d i o g r a p h i c studies of the peritoneal spread were done. Doses of isotopes were injected intraperitoneally. X-rays were given from 106 V a n d 250 kV machines.
Results Number. T a b l e 1 shows t h a t in the control g r o u p the average n u m b e r of mast cells per 20 fields was 310-4, This n u m b e r was increased in all the irradiated groups. R e m a r k a b l e increase was seen after irradiation with 1 0 6 V X-rays a n d Fe 59, n a m e l y 528 a n d 413 respectively. T h e average n u m b e r after t r e a t m e n t with ps2, Au 19s, a n d 250 kV X-rays did not show m u c h difference, the n u m b e r s being 344.7, 387, a n d 378.5 respectively. Size. T h e average area of cross section of the mast cells in control group was 4 2 3 . 8 / ~ . T h e rem a r k a b l e change was seen again for Fe ~9 with or without X-ray. Here most of the cells were very small, the average area being 151-1/~2 for Fe 59 a n d 140.3 #2 for Fe 59 along with 250 k V X-ray. T h e average area for p32 was 212.9/~ 2, for A u 19s 225.7 #2 a n d for b o t h 250 kV a n d 106 V X-rays 161-1 #2. Percentage of morphological variations. T h e various morphological characters observed were placed in 3 categories : 1. N o r m a l - - c e l l s with uniform distribution of m e t a c h r o m a t i c granules with or w i t h o u t visible nucleus. 2. Small h y p e r c h r o m a t i c - - c e l l s smaller t h a n the average of the above described n o r m a l cells a n d stained so deeply t h a t nuclei were not seen a n d the granules were found to be c l u m p e d together. 3. O t h e r s - - t h o s e which show g r a n u l a r shifting, bipolar giving a d u m b - b e l l shape, u n i p o l a r shifting of granules, small cells with nucleus filling almost the whole of the cells, cells with eccentric nucleus, cells with vacuolation a n d g r a n u l a r spilling, clumping of the granules in groups, cells h a v i n g peculiar shapes a n d cells showing degranulation. It is