Nikon L-ke User manual

Nikon
MICROSCOPE MODELL-Ke
INSTRUCTIONS
NIPPONKOGAKUK.K.
Copy offered by courtesy of Dr. Steven Horii, USA

Caution inHandlingandMaintenance
•Avoidtouchinglenssurfaceswiththefingersorwithanyroughmaterial.
Fordusting,usea featherora softlensbrushandthena washed-out
softcottonclothonlytowipeofffingermarksorgrease,wetthecloth
withxylor,butneverwithalcoholorether.
Themicroscopebodysurfacesshouldbedustedinthesamewayand
mayslightlybeoiled,
•Tightnessofthecoarsefocusingknobsshouldbeadjusted,inthistype
(L-Ke)ofmicroscope,bymeansoftheadjusting
ring, notbytwistingthe
knobs.
•Dismantlingoftheinternalopticalpartsandthemicroscopebodyshould
notbeattempted,becauseitmayinterferewiththeperformanceof
theinstrument.Itcanbeaccomplishedonlybyanexpertortheoriginal
manufacturer.
•Donotapplyanygreaseofnotspecifiedtypetotheslidingsurfacesof
thecoarsefocusingadjustmentandofthefloatingstage.Ifnecessary,
askforyourdealerorthemanufacturer.
•Themicroscopeistobeusedata dust-freeplace.Avoidanyforcible
manipulationofthemovingparts.Atalltimestheinstrumentistobe
handledcarefullye.g.forcarryingthemicroscope,holditsbasewithone
handanditsarmwiththeother.
•Whennotinuse,itshouldbecoveredwiththevinylcoverorkeptin
thewoodencabinetwhichisavailableonorder.Storeitata dryplace.
Itisrecommendedtokeeptheobjectivesandeyepiecesina desiccator.
NOMENCLATURE3
VARIOUSCOMBINATIONS5
CONTENTS 1-InterchangeableEyepieceTubes6
"U" Trinocular6
"T" Trinocular6
"B" Binocular6
"1" InclinedMonocular6
2. InterchangeableStages6
"R" RectangularMechanical6
"G" Graduated,CircularRotatable6
"C" CircularFloating6
"P" SquarePlain6
OBJECTIVES,EYEPIECES,CONDENSERS7
1. Objectives7
2. Eyepieces7
3.CombinationsofObjectivesandEyepieces8
4. Condensers9
ATTACHINGTHELENSES10
1. MountingtheObjectives10
2. MountingtheEyepieces11
3.MountingtheCondenser11
ILLUMINATION12
1. CondenserIrisDiaphragmandImages12
2. LightSource13
3.CondenserFocusingKnob13
4. BrightnessAdjustment14
5.PreparationandAdjustment14
6.Observation15
7.Photomicrography15
8.OtherLightSources15
9.FilterCase15
FOCUSING16
1. FocusingAdjustment16
2. EyepieceAdjustment16
3.CoarseFocusing17
4. PresetDevice17
5.FineFocusing•••18
6.OilImmersion18
7.LongWorkingRangeoftheStage19
8.ExchangingtheStage19
MOVINGTHESPECIMENONTHESTAGE19
1. RectangularMechanicalStage"R"19
2. CircularFloatingStage"C"20
EXCHANGINGTHEOBJECTIVENOSEPIECE••••21
PHOTOMICROGRAPHY••22

—3
NOMENCLATURE(MODELLBR-Ke)
Eyepiece
—4 —

VARIOUS COMBINATIONS
TheNikonMicroscopeModel L-Keis available in variouscombinations
with objectives,eyepieces,condensers,eyepiecetubes, and stages. Forex-
ample. Model LBR-Ke consists of Model L microscopestandwithKoehler
illuminator built-inbase,Binoculareyepiecetube and Rectangularmechani-
calstage.
Rectangular
Mechanical Circular
Floating Graduated,
Circular
Rotatable SquarePlain
— 5 —
1. Interchangeable Eyepiece Tubes
U=Trinocular
Magnificationfactor IX.Has provision for dioptercompensation and
interpupillarydistanceadjustmentfrom 54mm to 74mm. Observation
binocularsinclined45°,photo-tubeupright,360°rotatable.Withbuilt-in
sliding prismsystem,lighttransmission canbe switched over three
ways to permitphotomicrographythroughverticaltubewhileviewing
throughbinoculartube; 100% lightdirected to observationbinoculars
byswitching over lightpath; or wholelighttransmissiondirected to
verticalphototube for photomicrography,micro-projection or closed-
circuit T.V. pickup.
T—Trinocular
Magnificationfactor IX.Has fixedbeam-splitratio, 50%to observa-
tion binoculartubes and50%to verticalphotomicrographictube. In-
clined45° and rotatable 360°. Has provision for dioptercompensation.
Interpupillarydistanceadjustmentfrom 54mm to 74mm.
B = Binocular
Magnificationfactor IX. Inclined45° and rotatable 360°. Has provision
fordioptercompensation.Interpupillarydistanceadjustmentfrom
54mm to 74mm.
I = IndinedMonocular
Interchangeablewithothertypeeyepiecetubes.Inclined 45°and
rotatable 360°
2. Interchangeable Stages
R^RectangularMechanical
Stagesurface 130mm X 140mm. Has low-positionedcoaxial X andY
motion controlswhichprovideexceptionallyfine,smoothcrosstravel
within range of 50mmx75mm. Scales graduated to 0.1mm on vernier.
G = Graduated,CircularRotatable
Stagesurface 140mm in diameter.360°rotatable.Goniometerdivided
into 1°increments and reads to6' withvernier.Centerablestagepro-
videdwithclampingscrew.Suppliedwithstageclips.
C^Circuiar Floating
Stagesurface 140mm in diameterProvidedwithstageclips.Accepts
attachablemechanicalstageavailable on order.Movessmoothly in
anydirectionwithincirclediameter of 18mm in straightand/orrotat-
ing motion. Canbe clamped inany desiredposition.
P^Square Plain
Stagesurface 130mm X 130mm. Providedwithstageclips.Accepts
attachablemechanicalstageavailable on order.
- 6 -

OBJECTIVES, EYEPIECES, CONDENSERS
1. Objectives
a. OBJECTIVESformedical andbiologicaluse
Type Individual
Magnification NumericalFocal
ApertureLength Working
Distance Remarks
Achromatic Dry
Oil-immersion
4X "
lOX
20X
40X
S40X
NC40X
lOOX
SIOOX
lOOX
0.10
0.25
0.40
0.65
0.65
0.65
1.25
1.25
1.25
28.3mm
14.8mm
7.5mm
4.3mm
4.3mm
4.3mm
1.8mm
1.8mm
1.8mm
9.50lim
7.10mm
5.70mm
0.54mm
0.54mm
0.52mm
0.16mm
0.16mm
0.16mm
Spring-loaded
Nocoverglasstype
Spring-loaded
Withirisdiaphragm for dark-field
PlanAchromatic Dry
Oil-immersion
Plan 1.2X
2X
3X
4X
lOX
40X
NC40X
lOOX
0.03
0.05
0.08
0.10
0.25
0.65
0.65
1.30
35.8mm
42.3mm
37.7mm
29.5mm
15.6mm
4.0mm
4.1mm
1.6mm
29.7mm
35.6mm
28.6mm
18.2mm
7.0mm
0.24mm
0.32mm
0.12mm
Spring-loaded
Spring-loaded, no coverglasstype
Spring-loaded
PlanApochromatic
Oil-immersion
Apochromatic Dry
Oil-immersion
lOOX
40X
lOOX 0.80
„_1-40
1.6mm
4.3mm
L6mm
0.12mm
0.19 mm'
0.10mm
Spring-loaded
Spring-loaded
Spring-loaded
Long-WorkingDistanceType
Achromatic Dry LWD40X
4.0mm
2.0mm Fortissuecultureobservation
Phase-Contrast
Achromatic Dry
Oil-immersion
Long-WorkingDistance
Achromatic Dry
DLLIOX
B-MIOX
DLL20X
B-M20X
DLL40X
D-M40X
B-M40X
DLLIOOX
D-MIOOX
B-MIOOX
D-M40X
0.30
0.30
0.40
0.40
0.65
0.65
0.65
1.25
1.25
1.25
0.60
15.9mm
15.9mm
8.2mm
8.2mm
4.4mm
4.4mm
4.4mm
1.8mm
1.8mm
1.8mm
4.0mm
6.4mm
6.4mm
4.5mm
4.5mm
0.54mm
0.54mm
0.54mm
0.16mm
0.16mm
0.16mm
2.0mm
DarkContrast
BrightContrast
DarkContrast
BrightContrast
DarkContrast,Spring-loaded
DarkContrast //
BrightContrast
DarkContrast
DarkContrast //
BrightContrast n
DarkContrast if
(Fortissueculturework)
b.OBJECTIVESformetallurgicaluse
Metallurgy
Achromatic Dry
M5X
MlOX
M20X
M40X
MIOOX
0.10
0.25
0.40
0.65
1.25
25.0mm
14.8mm
7.5mm
4.3mm
1.8mm
15mm
7.1mm
5.7mm
0.52mm
0.16mm
PlanAchromatic Dry PlanMlOX
PlanM40X
PlanMIOOX
0.25
0.65
1.30
15.6mm
4.1mm
1.6mm
7.0mm
0.32mm
0.12mm
Spring-loaded
Spring-loaded
2. Eyepieces
Type Individual
Magnification Focal Length Field
Number Remarks
Higheyepoint,compensating HK5X 50mm 21.0 Withadjustableeyepiececollar
Higheyepoint,compensating,
wide-field HKWIOX 25mm 18.0 Withadjustableeyepiececollar
Higheyepoint,compensating,
wide-field HKW15X 16.7mm 14.0 Withadjustableeyepiececollar
Compensating,wide-field WFIOX 25mm 18.0 High-powerpurpose
Compensating K20X 12.5mm 8.0 Generalpurpose
Huygenian H5X
HlOX
H15X
50mm
25mm
^17nim
21.0
12.0
8.0
Generalpurpose
Generalpurpose
Generalpurpose
Diopteradjustable,high eye-
point,compensating,wide-field DHKWIOX 25mm 18.0 Suppliedtogetherwith
"U" eyepiecetube
With 5X,lOX,15X pictureframespluscross-
lines for framing and focusinsf
7 -
3. CombinationsofObjectivesand Eyepieces
Totalmagnifyingpowerobtainedbythecombinationistheproductof
individualobjectivepowermultipliedbyindividualeyepiecepower.A
selection ofthecombination will bedecidedsoastogetthehighest
resolutionoftheimage(resolvingpower),thelargestextentofobject
area (realfield)whichcanbeobservedwithoutmovingthestageor
slide, orthegreatestthicknessofobject(depthoffocus)whichcan
be distinctly seen withoutraisingorloweringthemicroscopestage,
depending uponthepurposeofusingthemicroscope.Shownbelow
are theresultscompiled from thedifferentcombinationsofobjectives
and eyepieces:
Objective Eyepiece Total
Magnifying
Power
Working
Distance
Resolution or
MinimumResolved
Distance RealField
ofView Depth
of
Focus
Objective Eyepiece Total
Magnifying
Power (mm) inobject
(^) inimage
(mm) (mm) (^)
4X 5X lOX
15X
20X
40X
60X 9.5
2.7-5.5
0.05-0.11
0.11-0.22
0.16-0.32
5.25
4.5
3.5
100
64
52
lOX 5X
lOX
15X
50X
lOOX
150X 7.1
1.1-2.2
0.05-0.11
0.11-0.22
0.17-0.33
2.1
1.8
1.4
16
10
8
20X 5X lOX
15X
lOOX
200X
300X 5.7
0.69-1.38 0.07-0.14
0.14-0.28
0.21-0.42
1.05
0.9
0.7
6
4
3
40X 5X lOX
15X
200X
400X
600X 0.54
0.42-0.84 0.08-0.17 0.17-0.34
0.25-0.50
0.52
0.45
0.35
1.8
1.2
1.0
lOOX 5X lOX
15X
500X
lOOOX
1500X 0.16
0.22-0.44
0.11-0.22
0.22-0.44
0.33-0.66
0.21
0.18
0.14
0.6
0.44
0.38
• Theworkingdistanceisthe clearance betweentheuppersurfaceofthe
cover
glass andthelowestedgeoftheobjectivecriticallyfocused.Note
that,asshownintheabovetable,theworkingdistancebecomesvery
small for high powerobjectives.
• Theresolutionofminimumresolveddistance(thelimitofresolvingpower)
is theminimumdistancebetweenobjectpointsdiscernibleasseparate
under themicroscopeilluminatedbythe light ofwavelength 550myu.
Theshorterthe light wavelength,thehighertheresolvingpower,that
is, thesmallresolvingdistanceresults.Inthetable,thesmallervalues
indicate theresolutionobtainedbyobliqueandthelargervaluesby
central illumination,(see"Illumination"onpage11)
#The minimumresolveddistanceintheimageisthevalueintheobject
multipliedbythetotalmagnificationofthemicroscope.Iftheresolving
powerofthemicroscopeisimportant,choosetheeyepiecebytheuse
ofwhichtheimageresolutioncomes
within thatofthenakedeye 0.15
—0.3mm (whentheobjectis seen from thedistance 25mm); generally
accepted criterion ontheupperlimitofthetotalmagnificationofa
-8-

microscopesetupisabout 500—lOOOX ofthenumericalapertureofthe
objectivetobeused.Notethatinphotomicrographyitisuselessto
raisethemagnificationbeyondtheresolvingpoweroftheemulsion
(usuallyabout 0.05mm). However, sincetheresolutionoftheemulsion
is higherthanthatofthenakedeye,photographsareusuallytakenat
alowermagnificationandthereaftertheyareenlarged.
• Realfieldofview(inmm)representstheextentoftheobjectthatcomes
underobservation.Inhighermagnificationitbecomesextremely small.
Consequently,itisadvisabletotakeaimattheobjectpointtobeexa-
mined firstunderlowermagnificationandthenrevolvethenosepieceto
highermagnification.
• Whenfocusisonthecenterofthe field, itscircumferencewillusuallybe
blurred, becausea curvatureoftheimageplaneisunavoidableinthe
microscope,exceptwhenusinga plantypeobjectives.Inordertoget
sharpcornerimage,itisnecessarytoadjustthefinefocusingknoband
switchthefocusfromthecenterontothecorners.
• Depthoffocusrepresentsthethicknessorheightoftheobjectinfi sharply
seenwhenobservationismadethroughthemicroscope.Inphotomicrogra-
phythedepthoffocusbecomessmallerthanthefigureshowninthe
previoustable.Therefore,carefulattentionmustbemadeinfocusing
when taking microscopepictures.
Byclosingthecondenserdiaphragm,thedepthoffieldcanbemade
greaterthanthevalueshowninthetable.
4. Condensers
Type
NumericalAperture Remarks
Abbe
1.30 Twolenses For centralillumination(withoutobliqueilluminationslider)
Abbe
1.30 Twolenses For centralandobliqueillumination(withoblique
illumi-
nation slider)
Aplanatic
1.40 Three lenses For high-classmicroscopy
Achromatic
1.25 Five lenses For high-classmicroscopy
Achromatic
(longfocal-lengthtype)
0.7 Four lenses Longfocal-lengthtype.Workingdistance12mm.With
turret-mounted annulardiaphragmsforphase-contrastand
phase-interferencemicroscopy.
UniversalDark-Field Condenser
1.20-1.40 for
dark-field
Suppliedincenterablemount.Withouterdiameter36.8
mm. ObjectivestobeusedlOXupto lOOX. Ideallysuited
for fluorescencework. lOOX objectiveusedshouldhave
built-in adjustableirisdiaphragm.Thicknessofslideglass
to beused,mustbelessthan 1.2mm.
Low-Power Condenser Singlelens For low-powermacro-objectives,e.g.,1.2X,2X,& 3X
PlanAchromatic
These condensersarenotonlycapableofconcentratinglight-beamforbetter
illumination oftheimage field, butalsogivesa greatinfluenceontheresolu-
tion ofmicroscopeimage,imagecontrastanddepthoffocus.Inprecise
observationandphotomicrography,theuseofanachromaticoraplanatic
condenserwiththemaximumnumericalaperture1.25providedwithan
obliqueilluminationdeviceanda filtercaseisspeciallyrecommended.
— 9 —
ATTACHING THELENSES
Beforeattachingtheobjectiveandtheeyepiecetothemicroscope,first
cleantheirouterlenssurfaces. Even a slightlystainedfingermarkmay
often interferewiththeimagecontrast.
1. Mounting the Objectives
Take specialcareinhandlingtheobjectives.
Beforeattachingtheobjectivesonthenosepiecerevolver,lowerthe
microscopestagesufficiently.Securingeachobjectivewiththefingers
ofonehand,screwit
into eachnosepiece
holewiththoseofthe
otherhand(Fig.4).
It isrecommendedto
mount theobjectives
onthenosepieceor-
derlyfromlowtohigh
powerssothatthe
magnification ofeach
objectiveaugmentsas
the revolverisrotated
clockwiseorcounter-
clockwise.
MicroscopeModelL-Kehasonitsuppersurfaceofthenosepiecerevolver
five blankspots(Fig.5)oneachofwhichtheobjectivemagnificationcan
beInkedso that, by
looking theseIndica-
tions fromabove,the
objectivelocatedbelow
canbeidentified with-
out havingtoturn
away your eyes from
the eyepiece.
Whenrotatingthere-
volver,holditsouter
milled rimwith your
thumb andfirstfinger,
but donotpushthe
objectivebarrels,other-
wisealignmentofthe
objectivesmaybe trou-
bled.
-10-

2. Mountingthe Eyepieces
Formounting,simplydrop the eyepieceinto the eyepiecetube. Itis
recommended to leave an eyepiece in placeevenwhen itisnotinuse,in
order to prevent the
entrance of dustinto
the eyepiecetube. Or
put onthe eyepiece
cap in place ofthe
eyepieceremoved.The
inclinedeyepiecebody
tube, trinocular,bino-
cular or monocular,can
berotatedafterloosen-
ing the clampscrew
(Fig. 6)for conveni-
ence in viewingfrom
anydesireddirection
without moving the
microscopestand. By furtherreleasing the clampscrew the bodytube
can be removed and replacedwithanothertypeeyepiecebodytube.
3. IVIounting the Condenser
To mount the condenser,unlock the lockscrew, and insert the condenser
beneaththe condenser
holder asfarasit will
go.Then,tighten the
lockscrew. In this
caselocate the dia-
phragmeccentering
lever andits screw at
aproperplace soas
to facilitatetheir
manipulation with one
hand (Fig.7).The
correctdistance tore-
tain immersion oilbe-
tween the lowersur-
face ofthe slide and
the topof condenser
issecuredwhen the
condenser is raised tothe upper limit by turning the condenserfocusing
knob.
-11—
ILLUMINATION
Resolution and contrast of image are greatlyaffected bythe method of
illumination.
1. Condenser IrisDiaphragmand Images
Stopdown the condenseririsdiaphragm and slide itinthe radialdirec-
tion fromcenter to edge. The farther the irisdiaphragm is off-centered, the
higherbecomes the contrast and resolution,whichdistinguishdetails of
object by increased and unsymmetricalshadow atthe boundary of object
(Fig. 8,a,b).
(a) Centralillumination(b)Obliqueillumination(c)Darkfieldillumination
When the irisdiaphragm is positioned soastoletthe lightbundleenterinto
the object atan angle o'f incidence the same asthe apertureangle ofthe
objective the resolutionreachesmaximum and twice as much asthe
resolution by centralillumination.
If the diaphragm is furthereccentered to such an extent asnotto
introduce the lightbundledirectlyinto the objective,darkfield illumi-
nationwill be obtained. Ifthe irisdiaphragm is widelyopened,images
byvariousangleilluminations are integrated. An illuminationangle not
favorable forthe object maybe included.
In the centralillumination the maximumresolution is obtainedwhen the
opening ofthe irisdiaphragmjustcorresponds tothe apertureangle of
the objective. In thiscase excessive outer rays tobe used as dark
field illumination arecutoffand flare is minimized. Ifthe opening is made
smaller, the contrast is enhanced,though the resolution is lowered. But
if the irisdiaphragm is largeenough to cover 60-70% ofthe objective
aperture, the decrease of resolutionwill notbe remarkable.
If the diaphragm is stoppeddown to minimum for admittingonly very
—12—

small lightbundles,theeffectsbydiffraction,reflection,refraction,etc.,
maybeexaggeratedsothatfringesmaybeseenattheimageedgeswhich
maylikelyinducemisinterpretationoftheimage,butitmaybeeffective
forspecialoccassions(e.g.definitionofgeneralstructureofnonstained
specimens).(Fig.8,c).
2. Light Source
Asalreadycited,theirisdiaphragmplaysanimportantroleInthe illumi-
nation onmicroscopy.Asa principlethediaphragmshouldbesoadjusted
that thenumericalapertureofthecondenserisequaltothatofthe
objectivebeingused,inordertoobtainthemaximumresolution.In
practice,however,tokeep
outstraylightwhichwould
reduceimagecontrast,
closingtheapertureofthe
condenserdownto60-
70% ofthatoftheobjec-
tivelenswillbringabout
agoodresultinmost
cases.Thecoincidence
ofcondenserdiaphragm
aperturewiththeopening
(exitpupil)oftheobjec-
tivecanbeascertained
bylookingthroughthe
microscopetubeafterre-
moving theeyepieceand
closingthediaphragm
slowly.Anexperienced
user,however,maydispensewithsucha procedure,andwillobtainthe
sameresultbyadjustingthediaphragmopeninguntilsatisfactorydistinct-
nessoftheimageisobtained.
Ifa highresolutionand,atthesame time, a highcontrastaredesirable,
obliqueilluminationwillbeeffective. This issuitedespeciallyforlowdyed
specimens,transparentphase-contrastspecimens,etc. However, inthis
illumination, a remarkablepolarityincontrastandresolutionmayappear;
it isnecessaryforobservationtochangethedirectionofillumination,by
turning theirisdiaphragm.
Thecondenserapertureisoff-centeredinanydirectionbyrotatingandat
the sametimebyradiallyslidingthediaphragm. This manipulationcanbe
doneonlybyusingonehand,thethumbandthefirstfingerforoff-centering
andthemiddlefingerforopeningorclosingthediaphragm.(SeeFig.9).
3. Condenser Focusing Knob
Thecondenserfocusingismadebyturningthecondenserfocusingknob.
This manipulationisnecessaryonlyincaseofKoehlerilluminationordark
-13-
field observation.Thecondenserisusuallytostayattheupper limit and
neednottobelowered,exceptwhena strayimageofanoutsideobject
superimposesthespecimenimagetoosharplyinsucha caseasindirect
sunlightilluminationorassomeunevenbrightnessinterfereswiththe
observation.
4. Brightness Adjustment
Forthispurposeadjustthevoltageofcurrentsupplybyregulatingthe
transformeroftheilluminatororbyusinga properneutraldensityfilter.
Adjustmentbymeansofthecondenserdiaphragmisnotatallpossible.
5. Preparation and Adjustment
(1)LampandSocket
AsshowninFig.10,lining
upthereddotonthesocket
retaining ringtothaton
the lamphousing,fitthe
sockettothehousingby
bayonetfashion.
(2)CenteringandFocusingof
the Lamp
Bringtheillumination
change-overleverinthe
position"M".Stopdown
the condenserdiaphragm
beneaththemicroscope
stage.Movethelampback
andforthtobringtheimage
ofthefilamentintosharp
focusonthebottomsur-
faceofthediaphragm.For
this purpose,itisconve-
nienttoutilizethereflection
onthe accessory reflecting
mirror, asshowninFig.11.Then,turnrightorleftonlythesocket
ring tofastenthelampsocketinposition.
Bringtheilluminationchange-overlevertotheposition"H".Manipu-
lating thelampcenteringscrews,makecenteringoftheimageonthe
condenserdiaphragm.
(3)CenteringoftheIlluminationFieldDiaphragm
Stopdowntheopeningoftheilluminationfielddiaphragmtoabout
2mm indiameter.
Viewingthroughthemicroscope,focustheobjective40Xtotheimage
ofspecimen.
Movingupanddownthecondenser,makea sharpfocusedimageof
the illuminationfielddiaphragmonthespecimen.
— 14—

Manipulatetwocenteringscrewsuntiltheimageoftheillumination
field diaphragmcomestothecenter.
Youneednotmindabouta slightdeviationfromthiscenteringcaused
byswitching over tootherobjectivesbecausethiswillnotproduce
anyobjectionableresult.
6. Observation
Forusualcaseofobservationwithobjectivesfrom4Xto lOOX (oil-immer-
sion),itsufficestoleavethechange-overleveratthepositionL.Whenan
exactilluminationorextremebrightnessisrequired,asisthecasewith
photomicrography,itisnecessarytomakeadjustmentforeachmagnifi-
cationbyswitching over thelever.
7. Photomicrography
Makeexactadjustment,dependinguponthemagnificationoftheobjec-
tivebeingused,byswitchingthechange-overleverasbelow:
Objectivemagnification4X
10-20X 40-lOOX
Positionofthechange-overleverL Mi H
i
Whenusinganobjective3Xorlower,setthechange-overlevertoH
andreplacethecondenserwiththeonespecificallydesignedforlower
magnification.
TheilluminationfortheModelL-Kewillnotchangeitscolortemper-
aturebythechange-overabovementioned.
8. Other Light Sources
Whenusinganyother
light sourceoutsideof
the microscope,replace
the illuminationfield
lenswiththe accessory
reflecting mirror,as
showninFig.12.
9. Filter Case
Afilterofthediameter 45mm canbemountedontheilluminationfield
—15—
lens.A filterofthediameter 33mm canbeplacedinthecondenseror
in thefiltercaseofthemicroscope. This filtercasetobeattachedonto
the microscopebase isavailableonorder.
FOCUSING
1. Focusing Adjustment
ThemicroscopemodelL-Keisprovidedwithcoaxial,coarseandfine
focusingknobs,bothofwhicharelocatednearthemicroscopebase(Fig.
13). Forwardrotationofeitherofthefocusingknobsbytheoperatorraises
the microscopestageandvice versa (Fig.13,14).
2. Eyepiece Adjustment
Whenusinga binocularortrinoculareyepiecetubeforobservationthe
adjustment oftheuser'seye-sight(diopter)discrepancybetweenthe
right andleft eyes isnecessarywhichismadebyrotatingtheadjusting
ring onthelefthandeyepiece.
Afterfocusingwith your righteyebyraisingorloweringthemicroscope
stage,turntheadjustingringleftorrighttoobtainthesharpimagewith
your lefteye,too.Then,regulatetheinterpupillarydistanceofthebino-
cularortrinoculartubebyslidingtheeyepiecesleftorrightbymeansof
the knob(Fig.15),untiltheviewfieldsofbotheyepiecesmergetogether.
It willbeadvantageoustomemorizetheattaineddiopterandinter-
pupillarydistancereadingsforfutureuse.
Thereddotengravedontheinterpupillarydistancescaleindicatesthe
positionwherethemechanicaltubelengthbecomesexactly 160mm. The
HK(higheyepointtype)eyepieceshaveaneyecupontop,therotationof
— 16—

whichwillgivepropereye-to-lensdistance. For thoseweareyeglasses,
eyecupshould be screwed in.
3. Coarse Focusing
Thecoarseadjustment maybe eased or tightened by means ofthe
adjusting ring:
If the revolution ofthe coarsefocusingknob istoo loose,turn thead-
justing ringcounterclockwise. Too muchtightness maybe adjusted bythe
clockwiseturning.
Never twist the focusingknobs for thisadjustment asinthe traditional
microscopewhosefocusingknobs,coarse and fine, are locatedseparate
(notcoaxial).Focusing maybe performed as follows:First,raise the
microscopestageuntil the distancebetween the specimen andthe
objectivebecomesless longerthan the workingdistance ofthe objective
to be used (see table onp.8), thenlookingthrough the eyepiece,lower
the stageuntil the specimen tobe examined is plainlyvisible.
4X, lOX,20X,40XandlOOX objectives are parfocal , andare approxi-
mately in focuswhenrevolvedintoposition one afteranother, theuse
of the finefocusingknobonlybeingrequired for criticalfocusing.
4. Preset Device
Theright-handfocusingknob hasa presetlever onits drum (Fig.16).
When the lever is fastened by turningclockwise (as indicated bythe arrow
engravedbesides)until it stops, the coarsefocusingknobscannot be turned
in the direction to drive the stagecloser tothe objective. This presetting is
— 17--
utilized for quickrefocusing
after thestage has beenlowered
andoutfocused for changing
the specimen or applying im-
mersionoil. The presetdevice,
whenfastened,prevents atthe
same time thedanger of damag-
ingthe objectivefront and slide
glass.
5. Fine Focusing
Manipulation ofthe finefocus-
ing knob is necessary:
a. To obtainthesharpestimage.
b. To transfer the focusfrom
center toa corner ofthe
viewfield.
c. To focusupon the differentlayers ofa thickspecimen.
d. To correct a slightblurringwhich may takeplacewhenshifting the
slide.
e.Tomeasure the thickness of objectunderexamination.
Themicroscope isso designedthat one revolution ofthe finefocusing
knob raises or lowers the microscopestage 0.2mm. This permitsdirect
reading onthe right-handknobscale upto 0.002mm (2«).The whole
range of finemovement is 38mm; the same asof coarsefocusing.
6. Oil Immersion
Whenusing lOOX objective, the application of immersion oilinthe
minute space(0.1mm)between the objective topandthe cover glass is
necessary to attain the specifiednumericalaperture. For criticalwork the
immersion oilistobe filledbetween thetop lens ofthe condenser and
the slide as well as between the objective andthe cover glass. Oilim-
mersionobservation is performed as follows:First,using lOXor40Xob-
jective (dry system),bring the specimen in focus andinthe center ofthe
viewfield. Setthe presetlever by turningclockwise. Lower the micro-
scopestage and revolve the nosepiecerevolver tolOOX objective.After
applying a drop of immersion oil onto the cover glass,raise the stage up
to the preset limit. Then,focus upby lookingthrough the eyepiece and
raising carefully the stage by manipulating the finefocusingknob. The
oil immersion lOOX objective is designed to attain its criticalfocusing by
about 1/3 forwardrotation ofthe finefocusingknob,that is, bringing
the stageabout 0.08mm closer tothe objectivefrom the parfocalposition.
Airbubbles inthe immersion oil, which may sometimesintervene the
microscopeimage andare visiblewhenlookinginto the microscope
tube without the eyepiece, canbe removed by repeatingslightmovement
of the nosepiecerevolver, by adding a certainquantity of immersion oil
or by means ofa needle.
Remaining stiffened oilmay oftenimpair the image.Therefore, im-
18

mediatelyafterfinishingthework,cleanofftheremainingoilfromthelens
usinga softcottonclothwettedwithxylor.Neverusealcoholorimmerse
thetopoftheobjectiveinxylor.
Becarefulnottouseimmersionoilthathasbeenthickenedbyage.
Therefractiveindexoftheimmersionoilshouldbe
1.515.
7.LongWorkingRangeoftheStage
ThemicroscopeL-Keis
designedtopermitlarge-
rangeloweringofitsstage
(71mm)forconvenience
inobservinga thickspeci-
mensuchasformetallurgy
orculture.Unlockthelock
screwbeneaththestage
(Fig. 17),andthestage
canbelowered33mm^^^^^^^
andbymanipulationof
^^W^^^^^gnm^^j^^^^ '[J
thecoarsefocusingknob
farther38mm.
Thestageloweredtosucha greatextentmaybeusefulinmacrophoto-
graphyorwhena specialstageisused.
8.ExchangingtheStage
Lowerthestagebymeansofthecoarsefocusingknobandthenunlock
thestagelockscrew.Thestage,then,canberemoved.
MOVINGTHESPECIMENONTHESTAGE
Clip
1. RectangularMechanicalStage'*R"
Thisstageenablesfinecrosswisetravellingoftheslideintherange
of50X 75mm,allowingreading
ofthemovementdownto0.1
mmbytheuseofthevernier
provided.
Forsecuringtheslideonthe
stageinposition,openthe
clip.
Eachtravellingisperformed
byrotationoftwocoaxialknobs
iDcated oneabovetheotheron
theverticalrodprotruded
belowontheleftsidefrom
thefront,theupperknobbeing
--19—
forlongitudinalandthelower
oneforlateraltravelofthe
slideonthestage.
Influorescencemicroscopy
orinusingoilimmersionob-
jectives,wheretheclearance
betweenthecondenserand
theslidealsoshouldbeoil-
immersed, stiffenedoilmay
causeunsmoothtravelofthe
slide. Inthiscase,removing
19ofthecircularopeningplate
atthecenterofthestageor
fasteningoftheclampscrew
willbehelpfultoa positive
traveloftheslide(Fig.18).
Byunlockingthelockknobon
theedgeofthestagethestage
canberotatedhorizontallyfor
convenienceintheobservation
fromtheoppositesideofthe
microscope(Fig.19),wherethe
eyepiecetubeistoberotated
180°. Thisrotationofthestage
mayoftenbeofuseinphoto-
micrography,whenthepicture
formatischangedfrom
verti-
caltohorizontalorviceversa.
Itisrecommendedtousethe
slideadapteronthestage
(Fig. 20)fora sufficientlongi-
tudinaltraveloftheslidein
suchreversedposition.
2. CircularFloatingStage
Thecircularfloatingstage
(Fig. 21)glidesandrotates
smoothlyandpreciselyinany
desireddirectionsimplyby
pushingtherimofthestage
bythefingers,wichina circle
of18mmindiameter.
Tofastenthefloatingstage
inposition,pressingthestage

22
downward, turn the rinn ofthe
stagecounterclockwise.
Fastening
ofthe floatingstage
isnecessary,whenusing
an
attachablemechanicalstage
(Fig. 22), whichwill be avail-
able
on order.
Thecircularrotatingstage
type
G is alsoavailable,which
permitsmeasurement
ofthe
rotatingangle of specimen
with
its circularscale.(Fig. 23).
EXCHANGING THE
OBJECTIVENOSEPIECE
It may becomenecessary
toexchange
the objective
nosepiece
in such a casewhen
aseries
of phase-contrast or
metallurgicalobjectives are
mounted asa unit onthe other
objectivenosepiece.
First,unlock
the revolver
lockscrew,
and draw outthe
nosepiece inthe direction ofthe
arrow (Fig.24).In place, in-
sert the othernosepiecegently
until
it stops,making the guid-
ingsurfaces ofthe dove-tail
fitparallel
to eachother.
Fasten
the revolver in position
bymeans
ofthe lockscrew
(Fig. 25).
-21—
PHOTOMICROGRAPHY
TheMicroscopeModel L-Ke, incorporating the Koehlerilluminationdevice
withlightsourcebuilt
inthe microscopebase,enablesconvenient and
perfectmicrophotographonly byadditionally mounting a cameraconnected
to
the microscopeeyepiecewith any photomicrographicadapter.
Therefore,whentakingphotographs
ofthe microscopicimage on 35mm
film, itis recommended tousethe NikonMicroflexModel EFM (withbuilt-in
exposuremeter)
orAFM (withbuilt-inexposuremeterpermittingautomatic
exposuresetting)
andthe Nikon F or Nikkormatcamera or NikonDark
Box
for usingwith the aboveattachments. For details of photomicrographic
methods,refer
tothe "Instructions of UsingNikonMicroflex EFM"or
"AFM" andany generalworks on photomicrography.
-22—

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