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LEITZ LABORLUX S Microscope for the examination of transparent objects

Instructions

WILD LEITZ

1 Care and Maintenance

Dust protection is provided by a flexible dust cover which shou ld al ways be u sed when the instrumen t is n ot in use. The stand sh ou ld be cleaned from t i me to ti me with a li nen or lea- t her cloth; a lcohol must not be used as i t attacks the paint, but pet roleum is well suited for cleaning tbe pa i nted surfaces.Pale spotson the object st age can be removed by rubbing wi th paraf- fi n oi l or vaseline. Particu lar care should be taken when undertaki ng studies using acids or ot her aggressive ch emicals. Di rect con tact of these substan ces with the stand or optics must be avoided u nd er a ll circu msta nces, and a H part s should be carefull y cleaned after use. The opti cs must be k.ept scru pu lously clean. Dust can be removed from glass surfaces by means of a d ry, fine-haired brush , blowi ng gently across the surface whilst brushing. Ifthe di rt is difficul t to remove, a clean cloth, mo is- tened with d ist i lled wa ter, can be used or, if this also has no effect, pu re alcohol may be applied . Particu lar care sh ou l d be taken w hen cleaning a n t i-reflection coatings. The ou ter eye- p iece surfaces and t he front elements of the object ives have coatings of ap proxi ma tely th e same h ardness asglass an d must be correspondi ngly carefu lly clea ned. Objectives shou ld not be screwed apart during cleaning. If damage or d i rt is noticed inside them, they shou ld be retu rned to us for repair. Cleanin g of the in ner su rfaces of the eyepieces is also advised against. Microscopes being used in hot a nd/or hu mid climates requ i re special ca re. It should be ensu red that a bu ild -up of fungu s does not occu r, wh ich is ma naged, i n the fi rst place, by tho- rough and m eticu lous cleaning a nd storage in a cu pboard whose i nside tempera ture is at least 5 °C above that of th e room. It m ust also be provi ded with airing h oles, loosely plugged wi th cotton wool or gauze as protec tion aga in st dust. If this type of storage is not possible, the microscope m ust be

kept in a closed contai ner with an adeq u a te amou n t of dry i ng agen t (e.g. silica gel). These measures should be t aken even in l aboratories wi th a i r cond i tioning. In warm and dry cl imates, dust is the greatest enemy. The i nst rumen t should, therefore, be covered wit h the d ust cover immed i ately after use or clea n- ing and stored in a cu pboard. Ifa humid period of longer tha n one month occurs, storage in a warm cupboard , as descri bed above, i s desirable. Proper handl ing of th e microscope will ensure decades of ser- vice. If, however, a check over or repa ir becomes necessary, please con tact you r Leitz agency or our Technica l Service direct. Technical Service Ernst Leitz Wetzlar GmbH, Postfach 20 07 D-6330 Wetzlar, West Germany. Telex: 483727 eltsc

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Fig. 1 Leitz LABORLUX S with stage no. 11, condenser no. 56 and binocular tube S. lEyepiece 2 Binocular Lube S 45° 3 Analyser sl it 4 Object ives 5 Specimen stage no. 11 6 Coax ial drive for moving the specimen 7 Screws to attach object guide no. 12 8 Height adjustment of the condenser mount 9 Condenser mou n t with slide changer

10 Adjustabl e height stop of the condenser m ount 11 Cen tri ng screws for condenser mou nt 12 Condenser no. 56 13 Field d iaphragm 14 Lamphousing 20 15 Mains switch (concealed)

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2 Assembling the microscope

Mounting the tubes Press the lever in t he direction of the arrow (Fig. 2); the t u be can now be i n serted into the q uick-change mount. Once the t u be has been i nserted allow the lever to slide back. The t u be can be rotated by 360° and clamped in a n y posi tion by slightly pulling the lever.

Fig. 2 Mou n t ing the tubes

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Inserting the eyepieces The eyepieces are i nserted i nto t he eyepiece t u bes (Fig. 3). For the LABORLU X S, Leitz eyepieces a re used which are calcu- lated for the mechanical tube length of 160 mm. These eye- pieces are distinguished from those for 170 mm tu be l ength by the addi tional identification of the field of view i ndex after the magnifi cat ion,e.g. lOx/20 . IfLeitz eyepieces wi thou! this iden- t ification of the field of view i ndex are to be used ,a spacer ring (ord er no. 519 653) must be push ed over the eyepiece mount on the side of the tu be.

Fig. 3 Inserting the eyepieces

The total magnification of the microscope is given by: reproduction ratio of the objective x eyepiece magnification (x tube factor). Example: 25/0.50 objective !Ox/18 eyepiece tube factor Ix Total magnificati on: 25 x 10 x 1= 250x

Screwing in the objectives Screw the object ives into the nosepiece (Fig. 4) in such a way that stepwise magnification chan ge is possi ble (e.g. in the order 4, 10, 40, etc). All Leitz microscope objectives from a magnification of 2.5x and calcul ated for a tube lengt h ofl60 mm ca n be used on the Leitz LABORLUX S. Please note in this cootext t11e field illu- mination capacity of the condensers. Microscope objectives calcul ated for 170 mm tube lengt h can be u sed from 16x magnification .

Fig. 4 Screwi ng i n the objectives

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Identification markings on the objectives (Fig. 5) 1 Mechanical lube length The distance in mm from the objecti ve should er Lo t he edge of the tu be . 2 Coverglass The engraving 0.17 js the thickness of the coverglass. A dash (-) i nstead of a number ind icates that the objective can be used for speci mens wi th or wit hout a covergl ass. 3 Field flatn ess of the objective EF objecti ves are systems with a more or less flat field of v iew of up to 18mm intermediate image diameter. PLAN objectives are systems with a flattened field of view up to 22.5 mm in termediate i mage diameter.

Fig. 5 Objective id en tification

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Ifthere is oo indication of field flattening, the objective is an achromat with optimu m correction, for use wi th u p to 18 field of view index.

4 Reproducti on scale in the intermediate image Size ratio between the intermedi ate image and the object, e.g. 10 :1

5 Numerical apert ure Physical id entifica tion of the objecti ve's resolv i ng power

6 [mmersion medium Immersion med ia can be, for example, oil,wa ter (W) or glycer- ine (G LYC) . The objective m ust always be used with the engraved immersion medi u m. Before the i mmersion objecti ve is focused, a drop of i mmer- sion medium should be applied to the object with out a ir bu bbles.

7 Phase contrast objectives Objectives wi th phase ring for phase contrast observa tion. These objectives have green wri ting (wi th the exception of achromats with black sleeves). The label for the condenser- side annu lar stops necessary for these objectives (e.g. PHACO 3) in a slide or in the turret pJa tc of t he UKL phase contrast condenser (e.g. PHACO 3 =set tu rret plate to 3) is a lso to be found on the phase contrast objectives.

8 Colou red ring indicating the objective magnification

9 Im mersion objectives

Attaching lhe object guide The object guide no. 12 for left- or right-hand operation is attached to the l eft or right of the specimen stage with two screws (6.1).

Inserting the condensers Rotate the cond enser clamp (2) un til the dots on the knurled knob and on the condenser mount coincide. Then lower the condenser using the knob (7.1) until the condensec (7.6) can be easily pushed i n to the mount as far as the stop. Now turn the condenser cla m p in the direct ion of t he arrow. The conden ser is n ow fim1ly fixed in the slide changer. Conden sers LK and U K.L are i nserted i n the same way as condenser no. 56 (illus- trated , 7.6).

Fig. 6 Attach ing the object gui de l Fixing screws for obJecL guide no. 12

Condenser no.56 Wit h the two-lens condern,er n o. 56, homogeneous illumina- tion i s posi-oi ble for all object fields ofobjectlves from 4x magni- fica tion at field of v iew index 20 withou t ad d itional swinging i n and ou t of the front lens. Condenser no. 56 workswith the n u me rical aperture A 0.90. lf oil cap no. 512 652 is screwed on, the numerica l aperture ca n be rai sed to A 1.25 when Leitz im mersion oil is a pplied between the condenser and t he speci men. To screw t he oil cap on, t he protective ri ng must be screwed off with a knurled ring.

Fig. 7 Inserti ng th e conden ser l Heigh t adj ustm ent of the conden ser h older 2 Condenser clam p 3 Condenser holder 4 Cent ring screws for condenser mount 5 Adjustable height stop for the con denser holder 6 Condenser no. 56

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The aperture diaphragm is adjusted with the lever. There is a scale for reproducible setting of the condenser diaph ragm. The sli t at th e side is i nten ded for the insert ion of sl ides wi t h annula r stops for sim pl e darkfield or phase contrast il lumina- tion. These can only be taken out of condenser no. 56 again after t he lever has been pressed.

Condenser LK The LK cond enser bas a h i nged condenser top S 1 .1. When the condenser t op isswung out ,the lower part of the conden6er can illu minate all object fields of obje ct ives from 2 .Sx upwards with field of view ind ex 20. When u sing objectives of lOx and higher the cond enser top m ust be swung i n .This is done wi th the hand le (8.2).

Fig. 7b Con denser n o. 56 l. Protective ring 2. Locking lever for 5lide with diffusion d isc and an nula r stops 3. Slit for slide w ith diffusion disc and annu lar stops 4. Lever for adjust ment of apertu re d iaphragm 5. Dovetail guide

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Wi th condenser top A 0.90 S I.I swu ng in, the cond enser LK works wi th the nu merical a perture A 0.90. Tl1e image of the field diaphragm is produ ced wit h t he conden ser top S l.l in a glass mediu m l.2 m m over t he stage surface. The condenser top of the LK cond enser is intercha ngeable v ia a thread wi th oth er condenser tops wi th a h igher nu merical aprtu re or lopswith a l onger working d istance. Darkfield con- denser tops D 0.80 or D 1.19 can also be screwed on instead of the S l.l standard top. Fu rther in formatio n on conden ser tops ca n be fou nd in t h e table on page 10. The apert ure iris diaphragm is adju sted with t he lever (8.3). There is a scale for reproducible sett ing of the condenser diaphragm.

Fig. 8 Condenser LK 1 A 0.90 S l.l Condenser top 2 Handle to swing ou t the condenser top 3 Lever for adjustment of the n perture diaphragm

Condenser UKL Its performan ce features in brigh tfield are the same as those of the cond enser no . 56. The annular slops necessary for phase contrast or darkiield illu- mination are situated in a turret plate (9.4) for quick change. Posit i on "H" (9.3) of the turret i s for brightfield (Hellfeld) investiga tions. Positions l , 2 and 3 accommodate the ann u lar stops PHACO l , 2 a n d 3. These can be aligned to th e phase ring of the objective in use by pushing i n the two centring screws (9.2). Position 4 is occupied by an ann ular st op for darkfield with object ives from lOx lo 40x. The wheel (9.5) is for adju sti ng t he apertu re d ia phragm , only necessary for brigh tfield investigati ons. It is fully opened for phase contrast or d arkfield (PH position).

f ig. 9 UK L condenser l Adjustmen t t elescope for centring the annular stops 2 Centring screws (press io to use) 3 Turret plate posit ion indication 4 Tu rret plate 5 Adjustment wh eel wi t h scale for aperture diaphragm

Darkfield condensers D 0.80 and D 1.19 The condenser D 0.80 is inten ded for darkfield investigations with lOx to 40x objectives. Image contrast is optimised here as compared with the darkfield produced with annu la r stops. The condenser D J.19 has an oil immersion cap and is used together with oil immersion objectives. The da rkfield cond en- ser caps are also avai lable withou t bottom parts and can be screwed onto the bottom part of the LK condenser. Fig. 10 Darkfield objective and funnel stop 1 Condenser top D 0.80 - 0.95 S 1.1 2 Oil condenser lop D 1.19 - 1.44 3 Lower pa.r t of conden ser with dovelai l gu ide 4 Special darkfield objective Ef 100/ 1.25 -0.60 Oel with built -in iris

diaph ragm 5 K nu rled r i ng for adjustmen t of the i ris diaphragm 6 Fu n nel stop for the oil i mmersion object i ve

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Condenser LK

UKL universal condenser for phase contrast

Condenser top

Revolver setting

Objective with engraving

Application

0.90 S 1.1 H All objectives Btightfield I PHACO l Phase contrast 2 PHAC0 2 Phase contrast 3 PHAC03 Phase contrast 5 All object ives Dark.field with aperture < 0.75

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Condenser Top

Top i n/out Use

0.90 S 1.1 Out With objective aperture <0.25 0.90 S 1.1 In With objective aperture >0.25 OEL 1.32 In

(Immersion oil on front element.) With OEL 100/1.32 objective

0.70 S4 In Intercept distance 4 mm. With specimen slid es of thickness >I mm.

0.55 S 15 I n Intercept distance 15mm . With specim en slides of thickness > 6 m m.

D 0.80-0.95 Cond enser top i n light path Darkfield. With objective apertures <0.75. D 1.19-1.44 Condenser top in light path

{Immersion oil on front element.) Darkfield. With objective apertu res < UO.

Attaching the lamphousing 20 Lamph ousi ng 20 is pl ugged i n to the guide sockets situated at the back of the stand a nd secured wi th the screw (1 1.1) (you may find it ha ndy to use a coin). Then connect the microscope via the connecting socket (11.2) wit h the mainscable lo the ma i ns. The switch (11.3) is for switchi ng the illumi na tion on and off. The lamp intensity ca n be regulated wit h t h e knurled knob (11.4).

Fig. J t Attaching l am phousi ng 20 I Fixing screw 2 Con necting socket for mains cable 3 On/off switch 4 I ntensity regulation

Changing the lamp Press the lamp socket (12.1), u nlock by turni n g to the left and tak e it out of the lamphou sing. Remove the d efect lamp.When i nserting t he new lamp,d o not rem ove its protect ive wrapp i ng until the lamp is in position. Return the lamp socket to t he lam- phousin g, press in and lock i n place by turni ng to the right.

Fig. U Changing the l amp I Lamp socket 2 I ntensity regulation

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Operation of the microscope

Focusing the image Place a specimen on t he stage or slide i t into the mounti ng plate (13.1) i f available. Specimen stage no. 11 (1.5) can be supplied with two cli ps to secure the specimen. I n stead of these cl ips,object guide no. 12 can be attached to the left or ri gh t of the stage for left- or ri gh t- handed operation (Fig. 6). It bas an adjustment range of 76 x 26 mm. Precise movement of the specimen within a range of 76x 52 mm is possi bl e with the mecha ni cal stage no. 18 (Fig. 13). The object guide no. 12 and mechanical stage no. 18 have a ver- nier scale (13.2) for adjustment of both the x and the y axis.

Fig. 13 Mechanical stage no. 18 I Speci men mou nting pl ate 2 Vern ier scale

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This enables certain parts of the specimen to be set reproduc- ibly. Tu rn in a mediu m-power object ive e.g. Ef'" 10/0.25 by rotating the nosepiece. Switch on the i llu mi nation (11.3) and regu late the intensi ty by turning k nob (12.2) . Turn the screw (7.5) by approx. 5 rota tions to the left and use the height adjustmen t (7.1) to m ove the condenser to the u pper st op nea r t he specimen. With the LK con denser, t he condenser top (8.1) is swung i n t o the illumi nating light path with the ha ndle (8.2). Ifyou chan ge to objectives of less than lOx magnification a fterwards, swing the cond enser top ou t of the i ll umi nat ing l i ght path. Open the apertu re diaphragm (8.3) and the field diaph ragm (1.13).

Adjustment of the binocular tubes I f u sing the bi nocular tu bes, adjust the in terpu pi lJ a ry distance u nt il the images for both eyes completely cover each other and appear as a single circular image. Then transfer the in terpu p il- lary distance(14.1) to t h escales(14.2) of the two eyepiece t ubes. How to compensate for vision defects: Look thro u gh the right-hand eyepiece with your right eye and focus the specimen with t he fi ne d rive. Then watch the same part of t be speci men with your left eye and rotate t he left-hand eyepiece tube until the samepa rt of the obje ct isin sharp focus. Do not use the fine d rive for this. This setting should be rechecked after central.ion of the condenser. When usi ng the binocula r FSA or FSA-R ph oto lu bes (Fi g. 14) the t u be length i s automatically compen sated when t he eye- pieces are set at the requ ired interpupillary d istance.

The sym bols on the rota ry knob of the bin ocular photo lubes Centring the condenser and setting the field diaphragm mean: Focus the specimen usi ng the coarse and fi ne drive.

100% light to t he eyepieces L 50 % l ight to the eyepieces,

50% to t he photo exi t l O 0Ai light to the eyepieces, 900/o to t he ph oto exit

The viewing angle of t he binocu lar SV and FSA-Y R tu bes can be adju sted by tilting the front part of the tu be between O and 40°. The u ser can th us set or alter the viewi ng height to su it himself.

Fig. 14 Selling the binocu lar L u bes

1. Close the field d iaphragm. Loosen tbe stop screw and raise the condenser to its u ppermost position with the height ad- j u stment (1.8).

2. Tu rn the cond enser stop screw (1.14) to lower the condenser until the edges of the field d i a p hragm appear sharp .

3. Centre the i mage of the field diaph ragm with the t wo cen- tering screws.

4. . Open t he fiel d d iaphragm until it ju sl disappears from the fiel d of v iew.

Fig. 15 Centering the condenser and setting the field diaphragm

II

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Use of the field diaphragm If optimum contrast is lo be allained, i t is important lo illumio- ale only tbe part of the specimen that appears in lhe image. Therefore the field diaphragm is only opened as far as the field of view. For this reason, a change of magoitica lion always necessitates matching the field diaphragm to the object field.

Use of the aperture diaphragm The more the aperture diaphragm is narrowed, t he higher the image contrast. Axial resolving power (image sharpness in the direct.ion of the optical axis) also increases. However, exces- sive closure of the aperture diaph ragm has an adverse effect on lateral resol v i ng power (selectivi ty). The best opt ical performance is obtained when the apert ures of the objective and the condenser are the same. A v isible deterioration of resolving power is seen when tbe apertu re diaphragm is closed over 1/3 of the aperture of the objective, and t bis should be avoided as far as possible. To check this, aneyepiece must be taken out of the eyepiece t u be, the apertu re diaphragm narrowed until i ts image covers 113 of the illumina ted rear lens area. Replace the eyepiece. If necessary, the apertu re diaphragm can be narrowed fu rther for objects with weak contrast. Once determ ined, the aperture diaphragm setting can be reprod uced with t he aid of the scales on the brightfield and phase contrast condensers. Note: The aperture diaphragm should not be used to set i mage inten- si ty. This is done with the rotary knob (12.2) for regulation of lamp intensi ty, or for photomicrography, wi th neutral l ight- blocki ng filters.

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Oil immersion objectives Oil immersioo objectives are labelled wi th the additional engraving "OEL" and a black ring on the lower edge of the objective mou nt. They must al ways be used wi th im mersion oil. Before the oil immersion objective is screwed into the nosepiece a d rop of immersion oil must be ap pl ied tothespeci- men. Check that the immersion oil is free from air bu bbles. The immersion oil has almost the same refractive index ne = 1.515 as the coverglass and the front lens of the microscope objective. The focal length and the working distance of ao immersion objective are usually very small. For this reason, worki ng wi th oil immersions req u i res great ca re. For rou tine work, the ord i nary 0.90 condenser is adeq uate even for oil immersi ons. However, if the fu l l apertu re of the im mer- sion objective is lo be u sed, e.g. for very fine structu res, the condenser apert ure must also be enlarged . For condenser no. 56 this can be done by screwing on a 1.25 OEL condenser cap, or for the LK condenser, an APL OEL 1.32 S I.I condenser top. In these cases, immersion oil must also be applied between the condenser cap/top and tbe underoeath of t he specimeo slide. After t he investigation, all surfaces where immersion oil has been applied must be carefu lly cleaned. Use a soft cloth mois- tened with alcohol or benzine.

Transmitted lighl dark.field with the LK condenser For darkfield investigations, condenser top D 0.80 -0.95 is used when the objective has an aperture of less than 0.75 and condenser top D 1.19 - l.44 when the objective'saperture islar- ger than 0.75. For apertures greater than L IO, insert a funnel stop into the oil immersion objective or use an objective with buill-in iris diaphragm. Separate bottom parts are also available for the darkfield condenser tops,which are pushed into the condenser mount i nstead of the brigbtfield or phase contrast condensers.

Focusing the darkfield image (D 1.19 -1.44 and D 0.80 - 0.95) Put a specimen on the stage. Turn the condenser stop screw to the right asfar as the stop. Insert thecondenser and raise lo the condenser stop. If u si ng the D 1.19 condenser top, first apply a drop of immer- sion oil to the surface of thecondense((laking care to avoid air bubbles) and then raise un til the drop of oil touches the under- neath of the specimen slide. This can be seen by a brief lighting u p of the slide. Bring the specimen into focus. (Use the 10/0.25 or 16/0.40 objective). Close the field diaphragm. Raise or lower lhe con- denser by adjusting the condenser stop screw (1.10) to the left and usin g the condenser drive (1.8) until the edgesof thedia ph- ragm are as sharp as possible when the specimen is observed . Move the image of the d i aphragm into the centre of thefield of view wi th the two centring screws (1.11). Open thefield diaphragm until it j ust disappears from the field of view. Phase contrast or darkfield illumination with condenser no. 56 Condenser no. 56 can be fitted with annular stop slides DF/ PHACO for phase contrast or darkfield illumi nat ion: - Centre the condenser witb a brightfield speci men - Screw in phase con trast objectives - Open the aperture diaphragm - Push the slide with the appropriate annular stop (chart on page 16) into the condenser (Fig. 16). The DF/55-56 annu lar stop slide is also available for conden- ser no. 56, with which simple dark.fiel d i llu mination can be obtained using object ives from !Ox to 40x.

Fig.16Insert ion of the slide with annular stop into condenser no. 56

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