Leica SP8 TCS User Manual
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1. Turn"the"PC"Microscope,"Scanner"Power,"Laser"Power,"and"the"Laser"Emission"key"to"on"
(bottom"right"of"desk)."
2. Turn"on"the"fluorescent"lamp"(top"left"of"the"computer"monitor)."
3. Log"into"the"computer"using"your"PI’s"last"name."
4. Double"click"on"the"LAS"X"software"found"on"the"desktop."
5. Select,"Configuration:"Maching.xhlw,"Microscope:"DMI8,"Load"Settings"at"Startup:"use"last"
system"settings"(This"should"be"default)."
6. Click"yes"if"you"want"to"use"image"stitching,"click"no"if"you"do"not"want"to"use"image"
stitching."
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63x"Plan"APO"1.4"NA"oil"immersion"
40x"HC"APO"1.3"NA"oil"immersion"
40x"HCX"PL"APO"0.85"NA"dry"
20x"HC"PL"APO"0.75"IMM"
10x"HC"PL"APO"0.4"NA"dry"
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The"objectives"are"selected"by"touching"the"desired"object"on"the"touch"screen."The"turret"will"turn"
automatically"to"the"selected"objective.""
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!"#$%&#%'()*+,)-./:"This"screen"highlights"th e "a p e rture"opening"and"th e"in t ensity"of"the"transmitte d "
light."TL=transmitted "ligh t "(2 )"to g g les "th e "sh u tt er "be tween"open"and"close d "fo r"b rig ht fie ld "
illumination."Intensity"and"Aperture,(3)"adjustment."(
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0"#1$%"#)#%2%$)*+,(F)G((Allows"the"user"to"switch"between"brightfield"(BF)"or"fluorescence"(FLUO)"(2)."
Do"not"touch"CS,or"POL."You"can"see"DAPI,"FITC,"o r"RHOD"through"the"eyep iec es "by "to u ch in g"th e "
appropriate"buttons"(3)."IL-Shutter,opens"and"closes"the"fluorescence"shutter"and"the"TL-shutter"(4)"
opens"and"closes"and"the"shutter"for"transmitted"light."You"will"only"see"the"IL-Shutter"when"you"
have"selected"fluoresce"and"you"will"only"see"he"TL-Shutter"when"you"have"selected"bright"field."
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The"SP8"is"an"inverted"microscope,"thus"the"objectives"are"below"the"specimen."Therefore,"if"using"a"
coverslip"slide,"you"must"turn"the"slide"upside"down."If"using"a"plate"it"must"be"a"specialized"glassbottom"dish,"with"a"thickness"of"1.5."When"using"an"oil"imme rsio n "o bje c tive "u se "on e "sm a ll"d ro p "of"o il,"
either"on"the"coverslip"or"the"oil"objective."HB(LB6(84M(BN1"&'(<C"(3%#(&#(E3<"#(;"'$"$O"Make"sure"the"
scan"head"is"tilted"all"the"way"forward,"otherwise"you"will"see"no"image"during"the"acquisition"phase.""
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To"move"the"stage"in"the"X/Y/Z"plane,"you"must"use"the"salt"and"pepper"shaker."The"top"knob"(Y)"moves"
the"stage"in"the"Y"plane"and"the"bottom "kno b"(X)"m o ve s"the"sta ge"in"th e"X"p lane ."You "can "togg le"
between"precise"and"fast"by"pressing"the"buttons"on"the"left"side"of"the"base."The"back"knob"controls"
the"Z"movement."You"can"toggle"b etw ee n"co arse "an d"fine "focu s"in"the"Z "plan e"by "using "the"bu tto ns"o n"
the"base"on"the"right"side."
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