1NTERPRETAHONa-OFULTRÀSTRUCTŒE
143
found these agent—s, as…well—as ethoxiÿethæol and r—hexyleneglyæl “to-have profound effects
on
the Size andb1refnngenæ of the mitotix apparatus. In Figure 18
‘shoWn a‘sz‘sula egg
111 metaphase of rst polar— body formation, the micrograph having been taken withre’ctied
objectives (Inoué -and Hyde,
If eggs—at this stage are
7—10% DMSO
‘
'
sea water, 10-15 % ethylene glycol in sea water or 2—3 % hexylene glyéol in sea Water (teni—
city being properly adjusted), the results :ate an increase
from-about ‘25'À
(measured between
to as much as 90 À, Figure l9.ThisinCreaSe
takes at most 5 minutes toæoccur and—is reversible if the * age‘ht fis ==Washed Out within
‘
10-15 minutes (depending upon the ‘stagewat Which itis applied)i'A'premmary analysis
with the electron microscope supports the Conclusions of Inoué'et al., 1965 forïa'simar
effect of D20, namely an increase in%the number Of
in the spindle,
Figure 20. An extensive analysis of this e"ect is:underway ad?Without—goihg into details we
suggest at this time that the primary e'ect of the glycols on
is to1 mobiliZe’ spindle
precursor material from a pOol of spindle material
theCell (Stevens, R.,
unpublished thesis, Inoué, et al., 1965) in‘to 'miCrotubules of the Spindleg- Further, the glycéls
'
do not affect isolated spindles at
else suggest that the glycols act ona control
the‘— proportion of
material between a precursor pool and mierotubules rathŒ!ïthà“à‘cühg£direétly
microtubules themselves.
‘
«
V
‘
-'
ï ‘ —
As has been pointed out in some detailin
which we consider to be due to-ice cr‘yStalS in
.
possibility of water removal due to
Conver—
sely, where we have deliberately removed
to that of chemicallyXed material as long asthawaterrreniovalïhas been in:?the-Ërange—‘of
40 to approximately 70 % (depending cri-tissue and stage)… This indicates to u‘s'ï(Withàll‘the
qualications necessary for interpretmgsa negative aràesult)*thätï morphology
to that
of chemically xed cells, can not'be obtained by eeiigÿte€hiciüëS une—sswœr:—is removed,
bound or changedin itsrelations
freezing protective agents is fm0st encouragmgthatav1abletechmquecanœsultfromrap1d
freezing work, since werhaveïseeh:almoçst3ïallà‘cellcomponentsseenmthesecellsbyusual
methods and indeed,
.
microvilli of the vitellinemembranegstructureswehavesænw1thnoothertechmquesWe
nofw have an additional source :‘of
consuder,however,thatappaæy due
to some formbf mbdiCati0n induced by: the freezing
cells ‘ « to
befrozen, resulting in
The
form of: modication is:t‘liat
PfÔ‘î
tective agents on spindles:ih
«
a large precursor pool of
canbemoblhzedfŒ
m1crotubule
'
formation. 011 the roman—hand
ay
;
'
,
great signicance
acüon °fffæzepmtæüveageS
investigating"the,Î'eçcts
fact,*âs
143
found these agent—s, as…well—as ethoxiÿethæol and r—hexyleneglyæl “to-have profound effects
on
the Size andb1refnngenæ of the mitotix apparatus. In Figure 18
‘shoWn a‘sz‘sula egg
111 metaphase of rst polar— body formation, the micrograph having been taken withre’ctied
objectives (Inoué -and Hyde,
If eggs—at this stage are
7—10% DMSO
‘
'
sea water, 10-15 % ethylene glycol in sea water or 2—3 % hexylene glyéol in sea Water (teni—
city being properly adjusted), the results :ate an increase
from-about ‘25'À
(measured between
to as much as 90 À, Figure l9.ThisinCreaSe
takes at most 5 minutes toæoccur and—is reversible if the * age‘ht fis ==Washed Out within
‘
10-15 minutes (depending upon the ‘stagewat Which itis applied)i'A'premmary analysis
with the electron microscope supports the Conclusions of Inoué'et al., 1965 forïa'simar
effect of D20, namely an increase in%the number Of
in the spindle,
Figure 20. An extensive analysis of this e"ect is:underway ad?Without—goihg into details we
suggest at this time that the primary e'ect of the glycols on
is to1 mobiliZe’ spindle
precursor material from a pOol of spindle material
theCell (Stevens, R.,
unpublished thesis, Inoué, et al., 1965) in‘to 'miCrotubules of the Spindleg- Further, the glycéls
'
do not affect isolated spindles at
else suggest that the glycols act ona control
the‘— proportion of
material between a precursor pool and mierotubules rathŒ!ïthà“à‘cühg£direétly
microtubules themselves.
‘
«
V
‘
-'
ï ‘ —
As has been pointed out in some detailin
which we consider to be due to-ice cr‘yStalS in
.
possibility of water removal due to
Conver—
sely, where we have deliberately removed
to that of chemicallyXed material as long asthawaterrreniovalïhas been in:?the-Ërange—‘of
40 to approximately 70 % (depending cri-tissue and stage)… This indicates to u‘s'ï(Withàll‘the
qualications necessary for interpretmgsa negative aràesult)*thätï morphology
to that
of chemically xed cells, can not'be obtained by eeiigÿte€hiciüëS une—sswœr:—is removed,
bound or changedin itsrelations
freezing protective agents is fm0st encouragmgthatav1abletechmquecanœsultfromrap1d
freezing work, since werhaveïseeh:almoçst3ïallà‘cellcomponentsseenmthesecellsbyusual
methods and indeed,
.
microvilli of the vitellinemembranegstructureswehavesænw1thnoothertechmquesWe
nofw have an additional source :‘of
consuder,however,thatappaæy due
to some formbf mbdiCati0n induced by: the freezing
cells ‘ « to
befrozen, resulting in
The
form of: modication is:t‘liat
PfÔ‘î
tective agents on spindles:ih
«
a large precursor pool of
canbemoblhzedfŒ
m1crotubule
'
formation. 011 the roman—hand
ay
;
'
,
great signicance
acüon °fffæzepmtæüveageS
investigating"the,Î'eçcts
fact,*âs