Book - Stoehr's Histology 2 - Revision history

Z8600021: /* Reconstructions */

2018-07-31T22:59:05Z

Reconstructions

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	drawing paper. The paper is on the table beside the base of the microscope.		drawing paper. The paper is on the table beside the base of the microscope.
	On looking through the camera into the microscope one can see the pencil		On looking through the camera into the microscope one can see the pencil
	point, as it is made to trace the outUne on the paper. In this way a succession of serial sections may be drawn with uniform magnification. The		point, as it is made to trace the outUne on the paper. In this way a succession of serial sections may be drawn with uniform magnification. The magnification is determined by substituting a stage micrometer for the shde
	~~magnificationis~~ determined by substituting a stage micrometer for the shde		of sections. The micrometer is a slide upon which 1 mm., with subdivisions into twentieths or hundredths, has been marked oflF by scratches in the glass; the subdivisions may be drawn with the camera under the same conditions as
	of sections. \ The micrometer is a slide upon which i mm., with subdivisions into twentieths or hundredths, has been marked oflF by scratches in the glass;		the sections, and the enlargement of the subdivisions may then be measured.
	the subdivisions may be drawn with the camera under the same conditions as
	the sections, and the enlargement of the subdivisions may then be measured~~^ â€¢~~

	From the camera-drawings of serial sections, wax reconstructions adult glands or embryonic organs may be made. If the sections are 10 fi
	thick and alternate sections have been drawn, magnified 50 diameters,		From the camera-drawings of serial sections, wax reconstructions adult glands or embryonic organs may be made. If the sections are 10 um thick and alternate sections have been drawn, magnified 50 diameters,
	then on the scale of the drawings these alternate sections are i mxn. apart.		then on the scale of the drawings these alternate sections are i mxn. apart.
	Wax plates imm. thick are therefore to be made, either by rolling the wax,		Wax plates imm. thick are therefore to be made, either by rolling the wax,
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	Often it is undesirable to attempt to make the magnification of the sections and of the side view identical; the measurements may be enlarged or reduced as they are transferred for plotting, by means of the		Often it is undesirable to attempt to make the magnification of the sections and of the side view identical; the measurements may be enlarged or reduced as they are transferred for plotting, by means of the
	draughtsman's proportional dividers~~^ â€”~~ an indispensable instrument forthis method of reconstruction. The corrections for unequal shrinkage of the sections in parajfin and other details can best be explained in the laboratory with the drawings at hand.		draughtsman's proportional dividers an indispensable instrument forthis method of reconstruction. The corrections for unequal shrinkage of the sections in parajfin and other details can best be explained in the laboratory with the drawings at hand.



	{{Lewis1906 footer}}		{{Lewis1906 footer}}

Z8600021: /* Special Stains */

2018-07-31T22:57:10Z

Special Stains

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Z8600021: /* General Stains */

2018-07-31T22:54:28Z

General Stains

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	==General Stains==		==General Stains==
			Haemaloxyline and eosine. Haematoxyline is a dye obtained from logwood, which stains nuclear structures blue. Eosine is an aniline dye staining protoplasm red. This and all aniline dyes used in histological stains should be prepared by Grubler in Germany.

	~~Haemaloxyline and eosine. Haematoxyline is a dye obtained from logwood,~~ which ~~stains nuclear structures blue. Eosine~~ is ~~an aniline dye staining protoplasm red~~. ~~This and all aniline dyes used in histological~~		There are many solutions of heamatoxyline among which is the following:
	~~stains should be prepared by Grubler in Germany~~.		* Haematoxyline crystals 1 gr.
			* Saturated aqueous solution of ammonia alum l00 cc.
			* Water 300 cc

	~~There are many solutions of heamatoxyline among which is the following:~~

	* Haematoxyline crystals 1 gr.
	* Saturated aqueous solution of ammonia alum l00 cc.
	* Water 300 cc

	Dissolve the crystals in the water, which may be heated, and add the alum		Dissolve the crystals in the water, which may be heated, and add the alum solution. Put the mixture in a bottle and drop in a crystal of thymol to prevent the growth of mould. A loose plug of cotton is used for a stopper and in this condition the solution is kept in the light for 10 days to ripen. It changes color during this process of oxidation, after which it is ready for use and is kept tightly stoppered. It deteriorates in a few months. If a strong solution is desired the amount of water may be reduced.
	solution. Put the mixture in a bottle and drop in a crystal of thymol to
	prevent the growth of mould. A loose plug of cotton is used for a stopper
	and in this condition the solution is kept in the light for 10 days to ripen.
	It changes color during this process of oxidation, after which it is ready for
	use and is kept tightly stoppered. It deteriorates in a few months. If a
	strong solution is desired the amount of water may be reduced.


	Another haematoxyline solution in common use is Delafield^s. It is		Another haematoxyline solution in common use is Delafield^s. It is made by dissolving 4 gr. of haematoxyline crystals in 25 cc. of 95 per cent, alcohol, and then adding 400 cc. of a saturated aqueous solution of ammonia alum. This is kept unstoppered for 3 or 4 days and then is filtered. 100 cc. each of 95 per cent, alcohol and of glycerine are added. It should not be used imtil it has become dark colored by remaining in the light for several days. Then it is to be filtered and tightly stoppered.
	made by dissolving 4 gr. of haematoxyline crystals in 25 cc. of 95 per cent,
	alcohol, and then adding 400 cc. of a saturated aqueous solution of ammonia
	alum. This is kept unstoppered for 3 or 4 days and then is filtered.
	100 cc. each of 95 per cent, alcohol and of glycerine are added. It should
	not be used imtil it has become dark colored by remaining in the light for
	several days. Then it is to be filtered and tightly stoppered.


	Eosine is sold in two forms, one soluble in water, the other in alcohol. In connection with the haematoxyline stain, a ~~yV ^^ i~~ per cent, aqueous solution may be used; or a i per cent, solution of alcoholic eosine, made in 60 per cent, alcohol.		Eosine is sold in two forms, one soluble in water, the other in alcohol. In connection with the haematoxyline stain, a ?? per cent, aqueous solution may be used; or a 1 per cent, solution of alcoholic eosine, made in 60 per cent, alcohol.


	To stain with haematoxyline and eosine the sections are placed in the haematoxyline solution from 2 minutes to an hour. They are then placed		To stain with haematoxyline and eosine the sections are placed in the haematoxyline solution from 2 minutes to an hour. They are then placed in water changed repeatedly for half an hour or longer (they may remain in it over night). As seen imder the microscope the nuclei should be deeply stained but the protoplasm should be nearly free from color. Stain in eosine for 1 to 5 minutes; dehydrate, clear, and moimt. For parafl&n sections this means treatment with 95 per cent., absolute and xylol, xylol, and damar. For celloidin sections, 95 per cent., oil of origanum, and damar.
	in water changed repeatedly for half an hour or longer (they may remain
	in it over night). As seen imder the microscope the nuclei should be deeply
	stained but the protoplasm should be nearly free from color. Stain in
	eosine for i to 5 minutes; dehydrate, clear, and moimt. For parafl&n
	sections this means treatment with 95 per cent., absolute and xylol, xylol,
	and damar. For celloidin sections, 95 per cent., oil of origanum, and
	damar.


	Methylene blue and eosine is highly recommended, especially for tissues		Methylene blue and eosine is highly recommended, especially for tissues fixed in Zenker's fluid and sectioned in paraflSn. Stain in a 5 or 10 per cent, aqueous solution of eosine for 20 minutes or longer, overstaining the tissue since the eosine is partly lost in the subsequent treatment. Wash out the excess of stain in water, and transfer to Unna's alkaline methylene blue diluted with three or four times as much water. Unna's blue is made by dissolving 1 gr. of methylene blue and 1 gr. of potassium carbonate in 100 cc. of water. Sections should be stained in the diluted solution for 10 to 15 minutes. Then they are washed in water and dehydrated and decolorized in 95 per cent, alcohol, moving the section about so that the stain may be washed out evenly. The pink color returns and when, as seen imder the microscope, the blue is limited to the nuclei the section is cleared in xylol and mounted in damar.
	fixed in Zenker's fluid and sectioned in paraflSn. Stain in a 5 or 10 per cent,
	aqueous solution of eosine for 20 minutes or longer, overstaining the tissue
	since the eosine is partly lost in the subsequent treatment. Wash out the
	excess of stain in water, and transfer to Unna's alkaline methylene blue
	diluted with three or four times as much water. Unna's blue is made by
	dissolving i gr. of methylene blue and i gr. of potassium carbonate in 100
	cc. of water. Sections should be stained in the diluted solution for 10 to
	15 minutes. Then they are washed in water and dehydrated and decolorized in 95 per cent, alcohol, moving the section about so that the stain may
	be washed out evenly. The pink color returns and when, as seen imder
	the microscope, the blue is limited to the nuclei the section is cleared in
	xylol and mounted in damar.


	Borax carmine and Lyons blue is perhaps the best general stain for		Borax carmine and Lyons blue is perhaps the best general stain for embryos. Dissolve 4 gr. of borax in 100 cc. of hot distilled water. When cool stir in 6 gr. of the best carmine and then add 100 cc. of 70 per cent, alcohol. After 24 hours, filter. The Lyons blue may be used in i per cent, alcoholic solution, made with 50 per cent, or 95 per cent, alcohol. Generally it is desirable to dilute it somewhat with alcohol before using.
	embryos. Dissolve 4 gr. of borax in 100 cc. of hot distilled water. When
	cool stir in 6 gr. of the best carmine and then add 100 cc. of 70 per cent,
	alcohol. After 24 hours, filter. The Lyons blue may be used in i per cent,
	alcoholic solution, made with 50 per cent, or 95 per cent, alcohol.
	Generally it is desirable to dilute it somewhat with alcohol before using.


	Before imbedding the tissue, it is stained in borax carmine from 24 to''		Before imbedding the tissue, it is stained in borax carmine from 24 to 48 hours, larger blocks of tissue requiring more time than small ones. After being placed in water for 5 minutes (a step which some omit), the tissue is transferred to acid alcohol (0.5 cc. of hydrochloric add in 100 cc. of 70 per cent, alcohol). In this the excess of stain comes out but the tissue acquires a deeper color. After remaining in the acid alcohol from 15 minutes to an hour the tissue is washed thoroughly in 70 per cent, alcohol and is imbedded and sectioned in parafiin in the ordinary way. After the sections have been attached to the slide they are stained in Lyons blue, rinsed in alcohol, dehydrated, cleared and mounted.
	48 hours, larger blocks of tissue requiring more time than small ones.
	After being placed in water for 5 minutes (a step which some omit), the
	tissue is transferred to acid alcohol (0.5 cc. of hydrochloric add in 100 cc.
	of 70 per cent, alcohol). In this the excess of stain comes out but the tissue
	acquires a deeper color. After remaining in the acid alcohol from 15
	minutes to an hour the tissue is washed thoroughly in 70 per cent, alcohol
	and is imbedded and sectioned in parafiin in the ordinary way. After the
	sections have been attached to the slide they are stained in Lyons blue,
	rinsed in alcohol, dehydrated, cleared and mounted.

	==Special Stains==		==Special Stains==

Z8600021: /* General Stains */

2018-07-31T22:52:44Z

General Stains

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	There are many solutions of heamatoxyline among which is the following:		There are many solutions of heamatoxyline among which is the following:

	Haematoxyline crystals i gr.		* Haematoxyline crystals 1 gr.
			* Saturated aqueous solution of ammonia alum l00 cc.
			* Water 300 cc

	~~Saturated aqueous solution of ammonia alum loo cc.~~

	~~Water *. 300 cc~~
	Dissolve the crystals in the water, which may be heated, and add the alum		Dissolve the crystals in the water, which may be heated, and add the alum
	solution. Put the mixture in a bottle and drop in a crystal of thymol to		solution. Put the mixture in a bottle and drop in a crystal of thymol to
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	and is imbedded and sectioned in parafiin in the ordinary way. After the		and is imbedded and sectioned in parafiin in the ordinary way. After the
	sections have been attached to the slide they are stained in Lyons blue,		sections have been attached to the slide they are stained in Lyons blue,
	rinsed in alcohol, dehydrated, cleared and mounted.		rinsed in alcohol, dehydrated, cleared and mounted.

	==Special Stains==		==Special Stains==

Z8600021: /* The Microscope */

2015-02-10T20:32:54Z

The Microscope

← Older revision		Revision as of 07:32, 11 February 2015
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	==The Microscope==		==The Microscope==

	It is trnfortimate that the price of a microscope is prohibitive to many medical students, and that some who would otherwise purchase instruments at the beginning of their work, wait until an official position entitles them to a discount. The price of microscopes is not always quite as high as is ~~Usted~~, and sometimes when several students buy microscopes at one time they may secure lower rates by having one of their number act as agent.		It is trnfortimate that the price of a microscope is prohibitive to many medical students, and that some who would otherwise purchase instruments at the beginning of their work, wait until an official position entitles them to a discount. The price of microscopes is not always quite as high as is listed, and sometimes when several students buy microscopes at one time they may secure lower rates by having one of their number act as agent.

Z8600021: /* The Microscope */

2015-02-08T09:00:59Z

The Microscope

← Older revision		Revision as of 20:00, 8 February 2015
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	==The Microscope==		==The Microscope==

	It is trnfortimate that the price of a microscope is prohibitive to many		It is trnfortimate that the price of a microscope is prohibitive to many medical students, and that some who would otherwise purchase instruments at the beginning of their work, wait until an official position entitles them to a discount. The price of microscopes is not always quite as high as is Usted, and sometimes when several students buy microscopes at one time they may secure lower rates by having one of their number act as agent.
	medical students, and that some who would otherwise purchase instruments at the beginning of their work, wait until an ~~ofl&cial~~ position entitles
	them to a discount. The price of microscopes is not always quite as high
	as is Usted, and sometimes when several students buy microscopes at one
	time they may secure lower rates by having one of their number act as agent.


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	Microscopes of a certain grade are required, and if they cannot be		Microscopes of a certain grade are required, and if they cannot be afforded, no instrument should be bought. The necessary equipment, as shown in the figure, is a stand with fine and coarse adjustment ("micrometer screw" and "rack and pinion'') and a large square stage. The more expensive round and mechanical stages are not necessary. There should be an Abb condenser (with iris diaphragm), a triple revolver, a high and a low eye-piece or ocular, and the following objectives: a f inch and a J or I inch, which must be parfocal, together with a iV oil immersion for
	afforded, no instrument should be bought. The necessary equipment, as		cytological and bacteriological work, and a 2 inch (very low power) for embryological work. The ^ oil immersion is an expensive objective, and its purchase may be postponed. The 2 inch is a cheap objective which is very useful in obtaining a view of an entire section, and for embryological reconstructions it is essential. The price of such an outfit, including the oil immersion objective, is from $70.00 to $90.00.
	shown in the figure, is a stand with fine and coarse adjustment ("micrometer screw" and "rack and pinion'') and a large square stage. The more
	expensive round and mechanical stages are not necessary. There should
	be an Abb^ condenser (with iris diaphragm), a triple revolver, a high and
	a low eye-piece or ocular, and the following objectives: a f inch and a
	J or I inch, which must be parfocal, together with a iV oil immersion for
	cytological and bacteriological work, and a 2 inch (very low power) for
	embryological work. The ^ oil immersion is an expensive objective,
	and its purchase may be postponed. The 2 inch is a cheap objective which
	is very useful in obtaining a view of an entire section, and for embryological
	reconstructions it is essential. The price of such an outfit, including the
	oil immersion objective, is from $70.00 to $90.00.


	Satisfactory microscopes of American manufacture are made by the		Satisfactory microscopes of American manufacture are made by the Bausch & Lomb Company. A sample submitted by the Spencer Lens Company to the Harvard Embryological Laboratory is also quite satisfactory. The Leitz microscopes, made in Germany, are preferred by some to the American instruments just described; they are not much more expensive. All agree that the Zeiss microscopes (German) are the best (and most expensive). It is undoubtedly true that any of these instruments will fill the requirements of medical students and physicians. If the microscope is purchased by a student unfamilar with its use, it is well to have the lenses examined by a disinterested microscopist.
	Bausch & Lomb Company. A sample submitted by the Spencer Lens
	Company to the Harvard Embryological Laboratory is also quite satisfactory. The Leitz microscopes, made in Germany, are preferred by some
	to the American instruments just described; they are not much more expensive. All agree that the Zeiss microscopes (German) are the best (and
	most expensive). It is undoubtedly true that any of these instruments will
	fill the requirements of medical students and physicians. If the microscope is purchased by a student unfamilar with its use, it is well to have the
	lenses examined by a disinterested microscopist.


	For a description of the nature and use of the microscope, the student		For a description of the nature and use of the microscope, the student is referred to the 9th edition of " The Microscope, " by Professor S. H. Gage, (Comstock Pub. Co., Ithaca, N. Y.).
	is referred to the 9th edition of " The Microscope, " by Professor S. H. Gage,
	(Comstock Pub. Co., Ithaca, N. Y.).


	For the sake of emphasis it may be said that the microscopist works		For the sake of emphasis it may be said that the microscopist works with his right hand upon the fine adjustment and his left hand upon the slide. As the latter is moved about, bringing diflFerent fields into view, the focussing is done with the adjustment and not with the eyes. Both eyes should be open (as will be natural after becoming, accustomed to the instrument). Often one acquires the habit of using only the right or the left eye for microscopic work, but it is better to learn to use both.
	with his right hand upon the fine adjustment and his left hand upon the
	slide. As the latter is moved about, bringing diflFerent fields into view,
	the focussing is done with the adjustment and not with the eyes. Both
	eyes should be open (as will be natural after becoming, accustomed to the
	instrument). Often one acquires the habit of using only the right or the
	left eye for microscopic work, but it is better to learn to use both.


	Always examine a specimen first with a low power and then with a		Always examine a specimen first with a low power and then with a high power objective. In focussing the microscope, have the objective drawn away from the slide and focus down. This should be done cautiously, with a portion of the specimen actually beneath the lens; if there is only cover glass and damar there, the objective will probably be driven down upon the slide. Unless one is sure that stained tissue is in the field, the slide should be moved back and forth as the objective is being lowered.
	high power objective. In focussing the microscope, have the objective
	drawn away from the slide and focus down. This should be done cautiously, with a portion of the specimen actually beneath the lens; if there is only
	cover glass and damar there, the objective will probably be driven down
	upon the slide. Unless one is sure that stained tissue is in the field, the
	slide should be moved back and forth as the objective is being lowered.


	In working with the Abb^ condenser the flat surface of the ~~minx)r~~		In working with the Abb^ condenser the flat surface of the mirror should be uppermost.
	should be uppermost.

	The objectives must never be scratched. Lens paper or fine linen		The objectives must never be scratched. Lens paper or fine linen should be used to wipe them. If they are soiled with damar they should be wiped with a cloth moistened with xylol. Since the lenses are mounted in balsam, xylol must be applied to them cautiously.
	should be used to wipe them. If they are soiled with damar they should
	be wiped with a cloth moistened with xylol. Since the lenses are mounted
	in balsam, xylol must be ~~appUed~~ to them cautiously.

	In lifting the microscope it should never be taken by any part above the stage; the pillar should be grasped below the stage.		In lifting the microscope it should never be taken by any part above the stage; the pillar should be grasped below the stage.

Z8600021: /* The Microscope */

2015-02-08T08:58:48Z

The Microscope

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	Within the past ten years the cost of a good instrument has been so reduced that an increasing proportion of students can enjoy the advantage of having a microscope of their own.		Within the past ten years the cost of a good instrument has been so reduced that an increasing proportion of students can enjoy the advantage of having a microscope of their own.

			[[File:Lewis1906 microscope.jpg\|600px]]


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	in balsam, xylol must be appUed to them cautiously.		in balsam, xylol must be appUed to them cautiously.

	In lifting the microscope it should never be taken by any part above the stage; the pillar should be grasped below the stage.		In lifting the microscope it should never be taken by any part above the stage; the pillar should be grasped below the stage.

	==Drawings==		==Drawings==

Z8600021 at 00:11, 6 February 2015

2015-02-06T00:11:05Z

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Z8600021 at 00:04, 6 February 2015

2015-02-06T00:04:25Z

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Z8600021 at 22:02, 4 February 2015

2015-02-04T22:02:26Z

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