Do you need a better microscope? Is research-grade your ultimate goal? Would you then be truly content? For me, the answers were always a wistful yes, yes and yes…

Then the daydream came true! Like all good stories, it's a tale of lessons learned with a happy ending and a promise of more to come. May I share it with you...?

My childhood interest in microscopy continued into adulthood where earnings financed an ongoing collection of prepared slides and the occasional microscope upgrade. Each upgrade improved on its predecessors in a fairly linear progression from toys through student models to small laboratory microscopes of mid 20^th century vintage.

Like most, I learned the craft by trial and error and by reading every microscopy book I could find. Many described techniques that my equipment didn’t support and this bred dissatisfaction with whatever microscope I owned at the time. It seemed the very books that inspired me also fuelled a nagging discontent by showing images captured with equipment I didn’t have. The wise say to squeeze the best from what you do own; sage advice indeed; but when you struggle to even detect amoebae in bright-field illumination it’s really hard not to covet phase contrast when you learn what it can do!

So far so stereotypical! Making-do with limited cash and equipment is a common feature of most amateur pursuits. There’s always better kit to lust after and I’m sure many of us dream of owning a fully-loaded research instrument. Yet we usually have to accept it is just a dream and we get by with whatever we can bodge, scrounge and (occasionally) afford instead.

But what if you could have a top-of-the-range research microscope stuffed to the hilt with top notch accessories? Assuming money was no object, would lasting contentment follow? Would you concentrate on the applications of your microscope instead of its improvement? I always wondered...

In December 2004 I was able to answer these questions myself. An opportunity to buy a Zeiss ICM 405 arose as a risky proposition “sold as seen” by an Ebay seller who claimed to “know nothing about microscopes”. The sale of my Victorian penny collection had left me with a very healthy hobby fund so after asking a few questions I put in a bid. Proximity to Christmas and negative wording must have scared off potential bidders because I won at the surprisingly “low” price of £1022. Would it arrive? Would it work if it did? Gulp...

Any worries were wholly unfounded as the scope arrived in perfect condition and loaded with every illumination technique I’d ever heard of and two that I hadn’t (Nomarski DIC and epi-fluorescence).The missus is very tolerant of my hobbies and I was allowed to set up temporarily at the kitchen table. The microscope is still there two years later! I claim to be the tolerant one now as I do allow the cooker, fridge, microwave and washing machine to be used in my lab!

My precious - still there today
 

Christmas and New Year passed in a pleasant blur as I discovered the capabilities of my new toy and learned to use them. The improvement in image quality over that delivered by all my previous scopes was astonishing. So much new detail was visible in my prepared slides it was like having an entirely new collection to explore. Oh, happy days!

I had secured a huge bargain and fulfilled a yearning for access to every technique I may ever wish to try. I learnt more about the limits of optical performance in three months of owning the Zeiss than I'd learnt in my entire life before. You’d think I’d be content now. Yes I was happy, ecstatic even, but the "equipment lust" only got worse!

The issue was that for the first time ever I actually saw the improvement in images delivered by a plan-apo objective over an achromat. If the condenser aperture was incorrectly adjusted or the lamp filament was misaligned, then the effect on image quality was as plain as day. High performance brought new sensitivity to limitations or maladjustments that were undetectable in my previous scopes (or if they were visible, I lacked the knowledge to recognise them). The boring optical theory I had skipped in those microscopy books was suddenly very interesting and I entered an intense period of re-reading and experiment to understand it better. It was most illuminating (ha-ha).

With my increasing (theoretical) understanding came an impression that even this fantastic scope could be improved. Use achromats? No chance! A research frame should be filled with plan-apos! Dark-field won’t do when ultra-dark-field is available. The condenser lens was N.A. 0.63 but my enthusiasm for working at the maximum resolution (fuelled largely by now owning a scope that supported it) demanded N.A. 1.4 too. More DIC sliders were needed because the scope had only two fitted. The film-based camera on the front photo port would have to go, so a digital SLR and imaging software topped the rapidly growing list.

As the basic scope was such a bargain, I felt it justified a generous budget to upgrade all of the accessories to top-of-the-line. Cash being readily available made the decision easy of course; perhaps too easy! I ring-fenced another £1000 (surely that would be enough) and began buying second-hand parts via Ebay and dealers.

Mmmm, parts... Drool

Mmmm, more parts... Slobber
 

One thing you learn when kitting out a research instrument is that this stuff is expensive! I mean really, really, really expensive! I'd been extremely lucky with the initial purchase. The £1000 approached £2000 before the refit was even near completion. Although mortally wounded, the hobby fund carried the project to its conclusion. Just.

Over the course of a year I equipped the scope for digital photomicrography, found more plan-apo objectives and added optional parts such as gliding stage, trinocular head, DIC sliders and optovar etc. I also got a halogen epi-illumination system and a set of Zeiss HD epi objectives to supplement the epi-fluorescence I rarely used (mercury lamp in a kitchen and all that). I made some mistakes buying incompatible or broken parts and I’ve yet to find a proper slide holder to replace my jury-rigged effort, but the system is all but complete. And what a system it is!

For £3500 I got a research microscope and accessories that sold for over £35,000 in 1980. It performs at the limits of optical resolution and is an ergonomic joy for imaging and visual use. It does bright-field, dark-field, ultra dark-field, POL, phase contrast, DIC, epi-fluorescence and epi-illumination (with BF, DF, POL and DIC on the epi path too). Any remaining limitations sit squarely with its owner.

So am I content? You bet! Extremely content! I feel free of any constraint and it’s a very nice feeling indeed. This scope is a keeper. To quote Homer Simpson, “woohoo”!

Did I overspend? Yes. Could I be content with less? No. It’s true that despite their exorbitant cost I seldom use the high-performance accessories now. What I thought was top priority has been tempered by practical experience. I do use them of course, but only when there's genuine need. Genuine need just doesn't turn up anywhere near as often as I thought it would. I think the real roots of my contentment lie in learning this for myself.

For example, using immersion oil was routine in the initial burst of interest but cleaning became a tiresome chore and my preference shifted to dry lenses when they resolved features I wanted to see. I bought objectives with correction collars for the benefit of dry usage, but they're tough to set up and suffer short working distances and appalling depth of field. High performance always demands compromise, complexity and crippling cost that's best avoided if you don't need it. But when you do need it, then nothing beats having the right gear to play with ;-) Check this out...

Pleurosigma Angulatum (LRGB composite)
 

Short wavelength monochromatic light is best for working near the limits of resolution, but by definition you can't get true colour images that way. This was an experiment to fix the problem in certain cases (e.g. nice iridescence that disappointingly vanishes on closer inspection). A very high resolution monochrome luminance image was scaled, aligned and combined with a normal RGB image for the L+RGB composite result above. Colour was a touch too (ahem) subtle in this specimen and I've yet to try it with others, but it looks a promising approach.

Fairly fiddly techniques went into the luminance channel and the result is mostly only interesting on a technical level. But high performance isn't always needed for pleasing images. If your goal is nice pictures, a dark-field stop and a 10x plan objective is one of many easier (and cheaper) ways to get equally satisfying results.

Portion of diatom circle – stack of 20 images combined in HeliconFocus
 

Most things I want to try now are supported at normal resolution with simpler techniques using the kit that first came with the scope. The original owner built a thoughtful package to a limited budget (ironically, much larger than mine as he bought it all new). It's the correct approach;

buy only what you need for what you want to achieve, and no more.

But I wanted to try everything before choosing what to achieve; particularly as the money was burning the proverbial hole. By following this advice I may have satisfied 95% of my needs at 1/3^rd of the cost, but that unavailable 5% would be a continuing source of discontent. And by amazing good fortune, despite my kid in a sweetshop approach, the system is still worth more than it cost. That's nice too.

So no regrets. It was hobby-money well spent and I can finally say "this is the last microscope I will ever buy". Really, it is. No. Really...!

It has lifted my interest and ambitions in microscopy to unprecedented levels, way beyond only browsing my slide collection and pond water. It offers such versatility and headroom that I’ve become a bit of a microscopical gadfly, flitting from one specialist field to another, just skimming the surface of most (for now). Despite 35 years fitful participation and vastly increased diligence over the past two years, I still haven't found my particular specialty. But I’m enjoying the search more than ever before. That'll do...!

Besides, for a proper specialty I'd probably need confocal laser imaging or something...

My scope doesn't do that!

Uh-oh...

All comments to the author Steve Beats are welcomed.

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